Determination of Hexosamines in Conjunction with Electrophoresis on Starch

Neuhaus, Otto W.; M, Letzring

doi:10.1021/ac60128a042

Cited by 130 publications

(44 citation statements)

References 21 publications

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“…An attempt was made to grade the degree of abnormality from the gross findings, as described previously (2). The Each precipitate was dissolved in 4 ml of water, and portions were used for determination of uronic acid with the carbazole method of Dische (9) and hexosamine by the method of Neuhaus and Letzring (10). Chondroitin sulfate chain length was determined by the procedure of Partridge, Davis, and Adair (11), which gives a number average molecular weight; reducing sugar determinations were done by the method of Somogyi (12) The in vitro synthesis of polysaccharide was studied using cartilage samples placed in ice immediately after removal at autopsy; finely diced samples weighing a total of 100 mg were incubated in Tyrode's solution containing 'SO4 or acetate-'4C, 5 ,uc per ml, at 370 C for 2i hours, then washed five times with 0.15 M saline and incubated overnight with trypsin as described above.…”

Section: Methodsmentioning

confidence: 99%

Biochemical Findings in Normal and Osteoarthritic Articular Cartilage. II. Chondroitin Sulfate Concentration and Chain Length, Water, and Ash Content*

Bollet¹,

Nance²

1966

J. Clin. Invest.

210

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Softening and erosion of cartilage, the characteristic early lesions of osteoarthritis, occur in sites of decreased chondroitin sulfate concentration. The role of aging and the specific biochemical events that produce this alteration in glycosaminoglycan composition have been under study in several laboratories. Collins and McElligott found increased radiosulfate fixation in these lesions compared with normal articular cartilage from the same individuals, suggesting increased synthesis of polysaccharide (1). These observations indicated an increased turnover of chondroitin sulfate in osteoarthritic cartilage lesions, and attention has become focused on the mechanism responsible for breakdown of this glycosaminoglycan.As reported previously (2), when the cartilage proteinpolysaccharide complex was extracted from human articular cartilage obtained at autopsy, a decrease in the concentration of chondroitin sulfate but not protein was found in the areas of erosion. This observation pointed to a specific breakdown of the glycosaminoglycan, since proteolysis might be expected to cause removal of both components such as occurs after papain injection into rabbits (3). An enzyme resembling testicular hyaluronidase could be responsible, but reports of the presence of hyaluronidase in tissues other than testis were unconvincing or unconfirmed. A reevaluation of the tissue distribution of this enzyme resulted in the finding of an enzyme with most of the characteristics of testicular hyaluronidase in

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Section: Methodsmentioning

confidence: 99%

Biochemical Findings in Normal and Osteoarthritic Articular Cartilage. II. Chondroitin Sulfate Concentration and Chain Length, Water, and Ash Content*

Bollet¹,

Nance²

1966

J. Clin. Invest.

210

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“…Gastric mucosal hexosamine was determined according to the method of Neuhaus and Letzring. 32) Briefly, gastric mucosal mucin was extracted with Triton X-100 and then hydrolyzed with hydrochloric acid. Hexosamine obtained from the hydrolyzed mucin was measured using acetylacetone reagent and Ehrlich's reagent.…”

Section: Methodsmentioning

confidence: 99%

Effect of Oral Vitamin E Administration on Acute Gastric Mucosal Lesion Progression in Rats Treated with Compound 48/80, a Mast Cell Degranulator

Ohta

Kobayashi

Imai

et al. 2006

Biological & Pharmaceutical Bulletin

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Compound 48/80 (C48/80) is known to cause degranulation of connective tissue mast cells, but not mucosal mast cells, with release of serotonin and histamine from the cells. 1,2) We have shown in rats with a single C48/80 treatment that the development of gastric mucosal lesions occurs with decreases in Se-glutathione peroxidase (Se-GSHpx) activity and vitamin E and hexosamine levels and increases in neutrophil infiltration, xanthine oxidase (XO) activity, and lipid peroxide level in the gastric mucosal tissue and that gastric mucosal blood flow changes like ischemia-reperfusion during gastric mucosal lesion development.3) We have also shown in rats treated once with C48/80 that neutrophils infiltrating into the gastric mucosal tissue participate in gastric mucosal lesion formation and progression, while the xanthine-XO system in the gastric mucosal tissue takes part mainly in the lesion progression. 4) Furthermore, it has been shown in rats treated once with C48/80 that acutely released endogenous serotonin contributes to gastric mucosal lesion formation, while released endogenous histamine mainly contributes to the lesion progression, although gastric acid plays no important role in the pathogenesis of the C48/80-induced gastric mucosal lesion.3,5) Ascorbic acid (vitamin C) is a water-soluble antioxidant and it scavenges reactive oxygen species (ROS), such as superoxide radical (O 2 Ϫ ), hydroxyl radical (OH . ), hydrogen peroxide (H 2 O 2 ), singlet oxygen, and hypochlorus acid, by itself and also supports the chainbreaking antioxidant action of vitamin E by reducing vitamin E radical to vitamin E at the liquid/aqueous interface. [6][7][8][9][10][11] In addition, ascorbic acid prevents neutrophil adherence to endothelium by scavenging ROS derived from activated neutrophils.12) Our recent reports have shown that ascorbic acid content decreases with a concomitant decrease in vitamin E content during lesion progression in the gastric mucosa of rats treated once with C48/80 and that gastric mucosal ascorbic acid plays a critical role in the progression of C48/80-induced gastric mucosal lesions, which is closely associated with its antioxidant and anti-inflammatory actions in the gastric mucosa. 13,14) Vitamin E is a lipid-soluble antioxidant and it functions as a chain-breaking antioxidant for lipid peroxidation in cell membranes and also as a scavenger of ROS such O 2 Ϫ , OH . , and singlet oxygen. 15) Vitamin E exerts an anti-inflammatory action by inhibiting the production O 2 Ϫ in activated neutrophils, adhesion of neutrophils to endothelial cells, and transendothelial migration of neutrophils. [16][17][18][19][20] Yoshikawa et al. 21) reported that both a decrease in gastric mucosal vitamin E level and an increase in gastric mucosal lipid peroxidation occurred during the development of ischemia-reperfusion-induced gastric mucosal injury in rats and that the severity of ischemia-reperfusion-induced gastric mucosal injury was enhanced in vitamin E-deficient rats. Naito et al. 22) have shown in nitric oxide-depl...

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“…Gastric mucosal hexosamine was assayed as follows: gastric mucosal mucin was extracted with Triton X-100 and then hydrolyzed with hydrochloric acid. Hexosamine obtained form the hydrolyzed mucin was assayed by the method of Neuhaus and Letzring 35) using acetylacetone, Ehrlich's reagent, and glucosamine as a standard.…”

Section: Methodsmentioning

confidence: 99%

Effect of Gefarnate on Acute Gastric Mucosal Lesion Progression in Rats Treated with Compound 48/80, a Mast Cell Degranulator, in Comparison with That of Teprenone

Ohta

Kobayashi

Imai

et al. 2005

Biological & Pharmaceutical Bulletin

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Gefarnate (geranyl farnesylacetate), an anti-ulcer drug, is clinically used for the treatment of gastric ulcers and gastritis.1,2) Takagi and Yano 3) reported that when gefarnate was orally administered to fasted rats for 5 d, the level of hexosamine, an index of gastric mucus, in the pyloric tissue of the stomach increased and that the reduction of hexosamine content in the gastric pyloric tissue of rats with 3 d of water immersion restraint stress (WIRS) was prevented by daily simultaneous administration of gefarnate. There are several reports showing that gefarnate exerts a protective effect against acute gastric mucosal lesions in various in vivo experimental models such as gastric mucosal lesions induced by immobilization, reserpine, fasting, ethanol, asprin, Hara et al. 6) showed that when pylorus-ligated rats with oral HCl-asprin treatment received a single oral pre-administration of gefarnate, this drug exerted a protective effect against gastric mucosal lesions with attenuation of decreased gastric mucosal hexosamine content. In addition, Kobayashi et al. 10) reported that gefarnate prevented WIRS-induced ulcer formation by inhibiting the reduction of endogenous prostaglandin E 2 and prostacyclin (prostaglandin I 2 ) levels.Teprenone (geranylgeranylacetone), of which chemical structure is similar to that of gefarnate, is an anti-ulcer drug developed in Japan. This drug is clinically used for the treatment of gastric ulcers and gastritis. 11,12) Teprenone is known to stimulate gastric mucus synthesis and secretion in rat gastric cultured cells [13][14][15] and in the gastric tissue of rats. 16,17) There are several reports showing that teprenone exerts a protective effect against acute gastric mucosal lesions in various in vivo experimental models, such as gastric mucosal lesions induced by asprin, cold restraint stress, WIRS, and HCl-taurocholate, through preservation of gastric mucus synthesis and secretion. 8,[18][19][20] It has been shown that teprenone protects cultured rat gastric mucosal cells against reactive oxygen species (ROS) such as superoxide radical (O 2 Ϫ ) by increasing the production of mucus. 21) In addition, it has been shown in vitro that teprenone inhibits the adhesion of neutrophils to endothelial cells and the expression of CD11b/CD18a, an adhesion molecule, on neutrophils when the neutrophils are activated by Helicobacter pylori water extract. 22) We reported that teprenone exerted protective and preventive effects against acute gastric mucosal lesions in rats with WIRS not only by preservation of gastric mucus synthesis and secretion but also by inhibition of neutrophil infiltration and enhanced lipid peroxidation in the gastric mucosa. 20)Compound 48/80 (C48/80) is known to cause degranulation of connective tissue mast cells, but not mucosal mast cells, with release of serotonin and histamine from the cells. 23,24) We have shown in rats with a single treatment of C48/80 that the development of gastric mucosal lesions occurs with decreases in Se-glutathione peroxidase acti...

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Determination of Hexosamines in Conjunction with Electrophoresis on Starch

Abstract: A modification of the Elson and Morgan procedure for glucosamine is readily applicable to establishing a "glycoprotein" pattern with zone electrophoresis on starch.

Cited by 130 publications

References 21 publications

Biochemical Findings in Normal and Osteoarthritic Articular Cartilage. II. Chondroitin Sulfate Concentration and Chain Length, Water, and Ash Content*

Biochemical Findings in Normal and Osteoarthritic Articular Cartilage. II. Chondroitin Sulfate Concentration and Chain Length, Water, and Ash Content*

Effect of Oral Vitamin E Administration on Acute Gastric Mucosal Lesion Progression in Rats Treated with Compound 48/80, a Mast Cell Degranulator

Effect of Gefarnate on Acute Gastric Mucosal Lesion Progression in Rats Treated with Compound 48/80, a Mast Cell Degranulator, in Comparison with That of Teprenone

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