scholarly journals Reaction of unsaturated uronic acid residues with mercuric salts. Cleavage of the hyaluronic acid disaccharide 2-acetamido-2-deoxy-3-O-(β-d-gluco-4-enepyranosyluronic acid)-d-glucose

1987 ◽  
Vol 245 (3) ◽  
pp. 795-804 ◽  
Author(s):  
U Ludwigs ◽  
A Elgavish ◽  
J D Esko ◽  
E Meezan ◽  
L Rodén
Keyword(s):  
Hyaluronic Acid ◽  
Connective Tissue ◽  
Uronic Acid ◽  
Glucuronic Acid ◽  
Reaction Products ◽  
Acid Moiety ◽  
Analytical Work ◽  
Almost All

Degradation of connective-tissue polysaccharides with bacterial or fungal eliminases and subsequent characterization of the reaction products are now part of standard methodology for the analysis of these compounds. However, the scope of preparative and analytical work based on the use of eliminases has been limited by the lack of procedures for specific removal of the unsaturated uronic acid residues generated in the eliminase reactions. In the present investigation, we have shown that these residues are cleaved by mercuric salts under mild conditions that are not likely to affect other structures in an oligo- or poly-saccharide molecule. Thus the disaccharide generated from hyaluronic acid by digestion with chondroitinase AC or ABC was cleaved into a keto acid and free N-acetylglucosamine within 10 min at room temperature upon exposure to 14 mM-mercuric acetate at pH 5. The reaction of the disaccharide with mercuric salts was used for ready determination of the distribution of radioactivity between the glucuronic acid and N-acetylglucosamine moieties in radioactive hyaluronic acid that had been synthesized by IMR-90 fibroblasts from 3H-labelled monosaccharides. When the precursor was [3H]galactose, over 95% of the incorporated radioactivity was found in the glucuronic acid moiety. In contrast, cells grown in the presence of [3H]glucosamine synthesized a polysaccharide in which almost all of the label was located in the N-acetylglucosamine units. It is apparent from these experiments that the reaction of unsaturated uronic acid residues with mercuric salts provides a new tool with potential for many applications in the study of the structure and metabolism of connective-tissue polysaccharides.

A uronic acid isomerase in Flavobacterium heparinum

1970 ◽  
Vol 48 (2) ◽  
pp. 164-169 ◽  
Author(s):  
James C. Karapally ◽  
Carl P. Dietrich
Keyword(s):  
Hyaluronic Acid ◽  
Equilibrium Point ◽  
Galacturonic Acid ◽  
Uronic Acid ◽  
Glucuronic Acid ◽  
Chondroitin Sulfates ◽  
Isolation And Characterization ◽  
Flavobacterium Heparinum

The isolation and characterization of a uronic acid isomerase from F. heparinum is described. The enzyme converts glucuronic acid and galacturonic acid to fructuronic acid and tagaturonic acid. The equilibrium point of the reaction is affected by buffers. In the presence of borate, 75% of glucuronic acid is converted to fructuronic acid. In the presence of phosphate, the equilibrium is reached when 31% of glucuronic acid is converted to fructuronic acid. Conversely, fructuronic acid and tagaturonic acid are converted to glucuronic acid and galacturonic acid, respectively. Monosaccharides derived from heparin and chondroitin sulfates do not affect the activity of the isomerase, in contrast to the monosaccharides from hyaluronic acid which have marked inhibitory (or diluting) activity upon the enzyme. The role of this enzyme in the metabolism of mucopolysaccharides is discussed.

THE SEPARATION, DETERMINATION, AND CHARACTERIZATION OF 2-AMINO-2-DEOXY-D-GLUCOSE (D-GLUCOSAMINE) AND 2-AMINO-2-DEOXY-D-GALACTOSE (D-GALACTOSAMINE) IN BIOLOGICAL MATERIALS BY GAS–LIQUID PARTITION CHROMATOGRAPHY

1964 ◽  
Vol 42 (4) ◽  
pp. 451-460 ◽  
Author(s):  
M. B. Perry
Keyword(s):  
Hyaluronic Acid ◽  
Chondroitin Sulphate ◽  
Chromatographic Determination ◽  
Partition Chromatography ◽  
Quantitative Analyses ◽  
Optical Rotations ◽  
Trimethylsilyl Derivatives ◽  
Quantitative Procedure

A method for the separation, determination, and characterization of 2-amino-2-deoxy-D-glucose (D-glucosamine) and 2-amino-2-deoxy-D-galactose (D-galactosamine) is presented. Treatment of 2-acetamido-2-deoxy-α-D-glucose and 2-acetamido-2-deoxy-α-D-galactose in pyridine solution with trimethylchlorosilane and hexamethyldisilazane results in a rapid conversion of the glycoses to their respective trimethylsilyl 3,4,6-tri-O-trimethylsilyl-2-acetamido-2-deoxy-α-D-glycosides which are sufficiently stable and volatile to allow their separation and quantitative analysis to be made by gas–liquid partition chromatography. The two trimethylsilyl derivatives, collected by preparative gas–liquid partition chromatography, were crystalline compounds which had sharp melting points and characteristic infrared spectra and specific optical rotations. Quantitative analyses of mixtures of 2-amino-2-deoxy-D-glucose hydrochloride and 2-amino-2-deoxy-D-galactose hydrochloride were made by gas chromatographic analysis of their trimethylsilyl derivatives formed after prior conversion to their N-acetyl derivatives.The analytical procedure was applied to the characterization of 2-amino-2-deoxy-D-glucose in hyaluronic acid and 2-amino-2-deoxy-D-galactose in chondroitin sulphate. The quantitative procedure was also successfully applied to the analysis of mixtures of hyaluronic acid and chrondroitin sulphate by the gas–liquid partition chromatographic determination of the 2-amino-2-deoxy-D-glucose and 2-amino-2-deoxy-D-galactose in the hydrolyzates prepared from synthetic mixtures of the two mucopolysaccharides.

The biosynthesis of the glucuronic acid moiety of hyaluronic acid

1953 ◽  
Vol 42 (2) ◽  
pp. 472-473 ◽  
Author(s):  
Saul Roseman ◽  
Julio Ludowieg ◽  
Frances Moses ◽  
Albert Dorfman
Keyword(s):  
Hyaluronic Acid ◽  
Glucuronic Acid ◽  
Acid Moiety

Compositional, microbiological, biochemical, volatile profile and sensory characterization of four Italian semi-hard goats' cheeses

2007 ◽  
Vol 74 (4) ◽  
pp. 468-477 ◽  
Author(s):  
Raffaella Di Cagno ◽  
R Evan Miracle ◽  
Maria De Angelis ◽  
Fabio Minervini ◽  
Carlo G Rizzello ◽  
...  
Keyword(s):  
Principal Component ◽  
Gas Chromatography Mass Spectrometry ◽  
Volatile Components ◽  
Volatile Profile ◽  
Mean Values ◽  
Trained Panel ◽  
Steam Distillation Extraction ◽  
Almost All

Four semi-hard Italian goats' milk cheeses, Flor di Capra (FC), Caprino di Cavalese (CC), Caprino di Valsassina (CV) and Capritilla (C), were compared for compositional, microbiological, biochemical, volatile profile and sensory characteristics. Mean values for the gross composition in part differed between cheeses. At the end of ripening, cheeses contained 7·98−8·51 log10 cfu/g of non-starter lactic acid bacteria. Lactobacillus paracasei, Lb. casei and Lb. plantarum were dominant in almost all cheeses. As shown by the Principal Component Analysis of RP-FPLC data for the pH 4·6-soluble fractions and by the determination of free amino acids, secondary proteolysis of CC and CV mainly differed from the other two cheeses. A total of 72 volatile components were identified by steam distillation-extraction followed by gas chromatography-mass spectrometry. Free fatty acids and esters qualitatively and quantitatively differentiated the profile of CV and CC, respectively. The lowest concentrations of volatile components characterized FC. Descriptive sensory analysis using 17 flavour attributes was carried out by a trained panel. Different flavour attributes distinguished the four goats' cheeses and relationships were found with volatile components, biochemical characteristics and technology.

Validation study of canine serum cortisol measurement with the Immulite 2000 Xpi cortisol immunoassay

2021 ◽  
pp. 104063872110292
Author(s):  
Jérémie Korchia ◽  
Kathleen P. Freeman
Keyword(s):  
Dynamic Testing ◽  
Total Error ◽  
Serum Cortisol ◽  
Cortisol Concentration ◽  
Canine Serum ◽  
Almost All ◽  
Serum Cortisol Concentration ◽  
Interpretation Of Results

We report the results of validation of canine serum cortisol determination with the Immulite 2000 Xpi cortisol immunoassay (Siemens), with characterization of precision (CV), accuracy (spiking-recovery [SR] bias), and observed total error (TEo = bias + 2CV) across the reportable range, specifically at the most common interpretation thresholds for dynamic testing. Imprecision increased at increasing rate with decreasing serum cortisol concentration and bias was low, resulting in increasing TEo with decreasing serum cortisol concentration. Inter-laboratory comparison study allowed for determination of range-based bias (RB) and average bias (AB). At 38.6 and 552 nmol/L (1.4 and 20 μg/dL), between-run CV was 10% and 7.5%, respectively, and TEo ~30% and ~20%, respectively (TEo remained similar regardless of the considered bias: SR, RB, or AB). These analytical performance parameters should be considered in the interpretation of results and for future expert consensus discussions to determine recommendations for allowable total error (TEa). Importantly, the commonly used thresholds for interpretation of results were determined ~40 y ago with different methods of measurements and computation, hence updating is desirable. Quality control material (QCM) had between-run imprecision of 4% for QCM1 and 7% for QCM2; the bias was minimal for both levels. Acceptable QC rules are heavily dependent on the desired TEa for the QCM system (TEaQCM), itself limited by the desired clinical TEa. At low TEaQCM (20–33%), almost no rules were acceptable, whereas at high TEaQCM (50%), almost all rules were acceptable; further investigation is needed to determine which TEaQCM can be guaranteed by simple QC rules.

scholarly journals EXPLORATION OF ORNAMENTAL FLORAS IN THE CAMPUS OF S.T. HINDU COLLEGE, NAGERCOIL, KANYAKIMARI DISTRICT, TAMILNADU, INDIA

2016 ◽  
Vol 3 (1) ◽  
pp. 59-64
Author(s):  
Mary Kensa V ◽  
Sahaya Anthony Xavier G ◽  
Asha A ◽  
Brintha B ◽  
Pechiammal M ◽  
...  
Keyword(s):  
Everyday Life ◽  
Public Spaces ◽  
Taxonomic Identification ◽  
Floristic Diversity ◽  
Floristic Survey ◽  
Observation Method ◽  
Almost All ◽  
Wild Progenitors

Most of the present day flowers have come from the wild progenitors, a few of which still exist in natural habitat.Ornamental flowers are highly promising and unutilized resources having tremendous and prover economic importance.Ornamental plants accompany people, since their birth to death and they coexist with almost all happy events in life such birthday celebrations, weddings, carrier progress etc. In addition, they form our best partners in our everyday life in our flats, offices, different public spaces, parks, gardens and elsewhere.An extensive floristic survey was conducted during the year 2015. Taxonomic identification, photographic documentation and ornamental characterization of each species with potential for use on floral art were recorded. The methodology used is based on observation method for the determination of flora. All the specimens collected were identified with the help of recent literature.The field expeditions of study area gave interesting results concerning floristic diversity.

A novel voltammetry offline coupled MALDI/TOF MS characterization of electrochemical reaction products and the voltammetric determination of febuxostat in human plasma

Talanta ◽  
2019 ◽  
Vol 194 ◽  
pp. 542-547 ◽  
Author(s):  
Fawzi A. El-Yazbi ◽  
Omayma A. Amin ◽  
Rania Bakry ◽  
Essam F. Khamis ◽  
Eman I. El-Kimary ◽  
...  
Keyword(s):  
Human Plasma ◽  
Electrochemical Reaction ◽  
Voltammetric Determination ◽  
Reaction Products ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Tof Ms

scholarly journals Specificity and action pattern of heparanase Bp, a β-glucuronidase from Burkholderia pseudomallei

Glycobiology ◽  
2019 ◽  
Vol 29 (8) ◽  
pp. 572-581 ◽  
Author(s):  
Yanlei Yu ◽  
Asher Williams ◽  
Xing Zhang ◽  
Li Fu ◽  
Ke Xia ◽  
...  
Keyword(s):  
Heparan Sulfate ◽  
Burkholderia Pseudomallei ◽  
Uronic Acid ◽  
Pathogenic Bacteria ◽  
Glucuronic Acid ◽  
Structural Determination ◽  
Action Pattern ◽  
Oligosaccharide Mapping ◽  
Size And Structure

AbstractThe specificity and action pattern of a β-glucuronidase derived from the pathogenic bacteria Burkholderia pseudomallei and expressed in Escherichia coli as a recombinant protein has been evaluated. While this enzyme shows activity on a number of glycosaminoglycans, our study has focused on its action on heparin, heparan sulfate and their biosynthetic intermediates as well as chemoenzymatically synthesized, structurally defined heparan sulfate oligosaccharides. These heparin/heparan sulfate (HP/HS) substrates examined varied in size and structure, but all contained an uronic acid (UA) residue β-(1→4) linked to a glucosamine residue. On the substrates tested, this enzyme (heparanase Bp) acted only on a glucuronic acid residue β-(1→4) linked to an N-acetylglucosamine, N-sulfoglucosamine or N-acetyl-6-O-sulfoglucosamine residue. A substrate was required to have a length of pentasaccharide or longer and heparanase Bp acted with a random endolytic action pattern on HP/HS. The specificity and glycohydrolase mechanism of action of heparanase Bp resembles mammalian heparanase and is complementary to the bacterial heparin lyases, which act through an eliminase mechanism on a glucosamine residue (1→4) linked to a UA residue, suggesting its utility as a tool for the structural determination of HP/HS as well as representing a possible model for the medically relevant mammalian heparanase. The utility heparanase Bp was demonstrated by the oligosaccharide mapping of heparin, which afforded resistant intact highly sulfated domains ranging from tetrasaccharide to >28-mer with a molecular weight >9000.

scholarly journals Characterization of structural component of cell walls of alkalophilic strain of Bacillus sp. C-125. Preparation of poly(γ-l-glutamate) from cell wall component

1987 ◽  
Vol 245 (2) ◽  
pp. 467-472 ◽  
Author(s):  
R Aono
Keyword(s):  
Cell Wall ◽  
Cell Walls ◽  
Glucuronic Acid ◽  
Molar Ratio ◽  
Gel Chromatography ◽  
Bacillus Sp ◽  
Cell Wall Component ◽  
Teichuronic Acid ◽  
Almost All

The cell wall of an alkalophilic strain of Bacillus sp. C-125 is composed of A1 gamma-peptidoglycan, a teichuronic acid and an unknown acidic polymer composed of glutamic acid and glucuronic acid, of which the molar ratio is approx. 4-5:1. Poly(gamma-L-glutamate) was prepared from the acidic polymer by removal of almost all of the glucuronic residues with trifluoromethanesulphonic acid treatment and purified chromatographically. The Mr of the polyglutamate preparation was estimated to be 14,000 by gel chromatography, or 43,000 on the basis of the content of N-terminal acid residues. The acidic polymer found in the cell wall of the organism was concluded to be a polyglutamate substituted with (oligo)glucuronic acid residues or a complex composed of two kinds of polymers (polyglutamate and polyglucuronate).

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