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Biomedical subjects

V L Murty

Publications and source records attributed to V L Murty.

At least 55 records · Page 3Linked to original sources
PubMed1990

Effect of lipids on the lactic acid retardation capacity of tooth enamel and cementum pellicles formed in vitro from saliva of caries-resistant and caries-susceptible human adults.

The lipid content and composition of these pellicles and the effect of lipids on their ability to retard the diffusion of lactic acid were investigated. Lipids accounted for 22.4% of the dry weight of caries-resistant enamel pellicle and 19.2% of caries-resistant cementum pellicle; the pellicle of caries-susceptible enamel contained 24.6% lipids and that of caries-susceptible cementum, 23.4%. Enamel and cementum pellicles from caries-resistant saliva had a significantly lower content of neutral lipids and phospholipids, whereas the glycolipid content was lower, although not significantly, in caries-susceptible pellicles. Pellicles from caries-resistant saliva had a considerably greater capacity to retard lactic acid diffusion than those from caries-susceptible saliva. In all cases, the retardation capacity was clearly dependent upon the lipid constituents, removal of which caused a 50-52% drop in lactic acid impedance by caries-resistant enamel and cementum pellicles, and 32-35% drop by caries-susceptible pellicles. On reacting the delipidated pellicles with their lipids, it was found that, in all cases, the highest quantitative effect on the restoration of the retardation capacity occurred when phospholipids were added. The findings suggest that the events controlling the interaction of salivary phospholipids with enamel and cementum may determine the susceptibility of the tooth surface to demineralization by acids produced by cariogenic micro-organisms.

Adult↗
PubMed1989

Physico-chemical characteristics of mucus glycoproteins and lipids of the human oral mucosal mucus coat in relation to caries susceptibility.

Mucus coat was isolated from oral epithelial surfaces of caries-resistant and caries-susceptible subjects, analysed for content and composition of lipids and mucus glycoproteins, and evaluated for physico-chemical characteristics. The mucus coat from caries-resistant subjects had a protein content similar to that of the caries-susceptible group but a higher content of carbohydrate and a lower content of lipids and covalently bound fatty acid. The carbohydrate component was mainly mucus glycoprotein, which accounted for 28.4% of the dry weight of caries-resistant mucus and 25.3% of caries-susceptible mucus. By chromatographic analysis on Bio-Gel A-50, both types of preparations had high (Mr approximately 2000 kdalton) and low (Mr approximately 300 kdalton) molecular-weight mucus glycoproteins. In the caries-susceptible mucus coat these two glycoproteins were in similar proportions, whereas the low molecular-weight glycoprotein predominated in caries-resistant mucus. In both preparations, the high molecular-weight glycoprotein was characterized by a high content of carbohydrates, associated lipids and covalently bound fatty acids, whereas the low molecular-weight glycoprotein was richer in protein and contained lesser amounts of associated and covalently bound lipids. Although the low molecular-weight glycoprotein showed only minor compositional differences with caries status, the high molecular-weight glycoprotein of the caries-resistant group had a 2.5 times lower content of covalently bound fatty acid, a 1.3 times lower content of associated lipids and contained 1.2 times more sulphate and sialic acid then that of the caries-susceptible group.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1989

Enzymatic sulfation of mucus glycoprotein in rat submandibular salivary gland.

1. The enzymic activity which catalyzes transfer of sulfate ester group from 3'-phosphoadenosine-5'-phosphosulfate to mucus glycoprotein was found associated with Golgi-rich membrane fraction of rat submandibular salivary gland. 2. Optimum enzyme activity was obtained with 0.5% Triton X-100, 4 mM MgCl2 and 25 mM NaF at a pH of 6.8 using desulfated submandibular salivary mucus glycoprotein. The apparent Km of the enzyme for mucus glycoprotein was 11.1 mg/ml. 3. Alkaline borohydride reductive cleavage of the synthesized 35S-labeled glycoprotein led to the liberation of the label into reduced oligosaccharides. A 75.4% of the label was found incorporated in four oligosaccharides. These were identified in order of abundance as sulfated penta-, tri-, hepta- and nonsaccharides. 4. Based on the results of chemical and enzymatic analyses of the intact and desulfated compounds the pentasaccharide was characterized as SO3H----GlcNAc beta----Gal beta----GlcNAc(NeuAc alpha----)GalNAc-ol and the trisaccharide as SO3H----GlcNAc beta----Gal beta----GalNAc-ol.

Animals↗
PubMed1989

Colloidal bismuth subcitrate (De-Nol) inhibits degradation of gastric mucus by Campylobacter pylori protease.

There is increased awareness that infection with Campylobacter pylori could be a major factor in the pathogenesis of gastric disease. Here, we present evidence that the extracellular protease elaborated by this bacteria, which causes degradation of gastric mucus, is inhibited by an antiulcer agent, colloidal bismuth subcitrate (CBS; De-Nol). The study was conducted with C. pylori cultured from antral mucosal biopsy specimens of patients undergoing gastroscopy. The grown colonies of bacteria were washed with saline, filtered through sterilization filter, dialyzed, and lyophilized. The powder was used as the enzyme source for proteolytic activity assay employing pig gastric mucus as substrate. Optimum enzymatic activity was obtained at 37 degrees C and at pH 7.0. The apparent Km of C. pylori protease with gastric mucus was 0.71 g/L. Analyses of the degradation products indicated that the protease caused extensive proteolysis of mucus glycoprotein polymer. Introduction of CBS to the incubation mixtures led to a reduction of the rate of mucus degradation. The rate of proteolysis inhibition was proportional to CBS concentration up to 1 X 10(-1) g/L, at which point a 37% reduction in mucus proteolysis was obtained. The Km value for proteolytic degradation of mucus by C. pylori protease in the presence of CBS was 1.25 g/L. The results suggest that CBS is capable of counter-acting the proteolysis of the protective gastric mucus layer by C. pylori.

Animals↗
PubMed1989

Effect of antiulcer agents on the physicochemical properties of gastric mucus.

Although the mechanism of gastric mucosal protection is multicomponential, the initial brunt of luminal insult falls on the layer of mucus weakening its physicochemical characteristics and is intimately associated with gastric disease. Hence, the agents capable of strengthening the qualities of mucus gel, associated with its protective function, are becoming increasingly popular in peptic ulcer therapy. Evaluation of the efficacy of these types of drugs requires delineation of their direct effect on the mucus layer constituents, from those evoked in the chemical composition of mucus as a result of prolonged drug administration. Studies with mucus layer strengthening agents indicate that such drugs as sucralfate and De-Nol interact tenaciously with the mucus gel and in essence seal the underlying mucosal surface from the noxious luminal contents. These drugs and other new antiulcer agents like sofalcone, geranylgeranylacetone and stable prostaglandin analogs are also capable of enhancing mucus gel viscosity and its hydrogen ion impedance ability. Furthermore, the majority of these drugs exert an inhibitory effect on peptic degradation of gastric mucus, and the colloidal bismuth preparation (De-Nol) is also capable of prolongation of the luminal availability of epidermal growth factor. The data on gastric mucus gel, following prolonged administration, show that these drugs enhance the lipid content of the mucus gel, and increase both the mucus coat dimension and the proportion of the high molecular weight form of its mucin component. These changes result in the improvement of viscoelastic, permselective and hydrophobic properties of gastric mucus. Hence, continuity of the function of mucus layer perimeter of mucosal defense is maintained.

Anti-Ulcer Agents↗
PubMed1989

Campylobacter pylori colonization factor shows specificity for lactosylceramide sulfate and GM3 ganglioside.

The specificity of Campylobacter pylori cell surface lectin, a presumptive colonization factor, was investigated using various sulfated and sialic acid containing glycolipids. C. pylori cells, cultured from human antral mucosal biopsies, were incubated with intact and modified glycolipid preparations and examined for agglutination inhibition of human erythrocytes. Titration data revealed that the inhibitory activity was highest with lactosylceramide sulfate and GM3 ganglioside, while galactosylceramide sulfate GM1, GD1a and GD1b gangliosides were less effective. A strong inhibitory activity towards C. pylori hemagglutin was also observed with an antiulcer agent, sucralfate. The inhibitory effect of both types of glycolipids was abolished by the removal of sialic acid and sulfate ester groups, thus indicating that sulfated and sialic acid containing glycolipids with terminal lactosyl moieties serve as mucosal receptors for colonization of gastric epithelium by C. pylori.

Animals↗
PubMed1988

Sulfation in vitro of mucus glycoprotein by submandibular salivary gland: effects of prostaglandin and acetylsalicylic acid.

Enzymatic sulfation of mucus glycoprotein by rat submandibular salivary gland and the effect of prostaglandin and acetylsalicylic acid on this process were investigated in vitro. The sulfotransferase enzyme which catalyzes the transfer of sulfate ester group from 3'-phosphoadenosine-5'-phosphosulfate to submandibular gland mucus glycoprotein has been located in the detergent extracts of Golgi-rich membrane fraction of the gland. Optimum enzyme activity was obtained at pH 6.8 with 0.5% Triton X-100, 25 mM NaF and 4 mM MgCl2, using the desulfated glycoprotein. The enzyme was also capable of sulfation of the intact mucus glycoprotein, but the acceptor capacity of such glycoprotein was 68% lower. The apparent Km of the submandibular gland sulfotransferase for salivary mucus glycoprotein was 11.1 microM. The 35S-labeled glycoprotein product of the enzyme reaction gave in CsCl density gradient a 35S-labeled peak which coincided with that of the glycoprotein. This glycoprotein upon reductive beta-elimination yielded several acidic 35S-labeled oligosaccharide alditols which accounted for 75% of the 35S-labeled glycoprotein label. Based on the analytical data, the two most abundant oligosaccharides were identified as sulfated tri- and pentasaccharides. The submandibular gland sulfotransferase activity was stimulated by 16,16-dimethyl prostaglandin E2 and inhibited by acetylsalicylic acid. The rate of enhancement of the glycoprotein sulfation was proportional to the concentration of prostaglandin up to 2.10(-5) M, at which point a 31% increase in sulfation was attained. The inhibition of the glycoprotein sulfation by acetylsalicylic acid was proportional to the drug concentration up to 2.5.10(-4) M at which concentration a 48% reduction in the sulfotransferase activity occurred. The apparent Ki value for sulfation of salivary mucus glycoprotein in presence of acetylsalicylic acid was 58.9 microM. The results suggest that prostaglandins may play a role in salivary mucin sulfation and that this process is sensitive to such nonsteroidal anti-inflammatory agents as acetylsalicylic acid.

16,16-Dimethylprostaglandin E2↗
PubMed1988

Prostaglandin effect on the enzymatic sulfation of mucus glycoprotein in gastric mucosa.

The effect of 16,16-dimethyl prostaglandin E2 (DMPGE2) on the sulfation of mucus glycoprotein in gastric mucosa was investigated. The enzymatic activity which catalyzes the transfer of the sulfate ester group from 3'-phosphoadenosine-5'-phosphosulfate to gastric mucus glycoprotein was located in the detergent extracts of Golgi-rich membrane fraction of antral and body mucosa of rat stomach. The sulfotransferase activity of this fraction from body mucosa, however, was 35% higher than that from the antrum. The enzyme exhibited optimum activity at pH 6.8 using 0.5% Triton X-100 and 30 mM NaF. The apparent Km of the enzyme for sulfation of mucus glycoprotein was 10.5 microM, and the sulfate ester was found incorporated into the carbohydrate chains of the glycoprotein. Introduction of DMPGE2 to the reaction mixtures led to an enhancement in the rate of mucus glycoprotein sulfation. The rate of enhancement was proportional to the concentration of DMPGE2 up to 1.0 x 10(-4) M and was of the competitive type, with an apparent Km value of 6.7 microM. Since sulfated mucins play an important role in gastric mucosal defense and the increase in their sulfation occurred at levels of prostaglandin present in gastric mucosa, the observed effect may be of significance to gastric mucosal defense in vivo.

16,16-Dimethylprostaglandin E2↗
PubMed1988

Role of salivary epidermal growth factor in the maintenance of physicochemical characteristics of oral and gastric mucosal mucus coat.

The involvement of salivary epidermal growth factor (EGF) in the maintenance of oral and gastric mucosal mucus coat dimension and chemical characteristics was investigated using sialoadenectomized rats. Examination of the oral and gastric mucosal surface by phase contrast microscopy and Alcian blue uptake revealed that deprivation of salivary EGF caused a 31-36% reduction in mucus coat thickness and a 38-43% reduction in adherent mucin content. Chemical analyses indicated that the mucus coat of sialoadenectomized group exhibited a 21-28% increase in protein and a 67% decrease in covalently bound fatty acids, a 30% decrease in carbohydrates, and a 32-37% decrease in lipids. Sialoadenectomy also evoked changes in the chemical composition of mucus glycoprotein component of oral and gastric mucus coat reflected in the lower content of sulfate (25-26%), associated lipids (24-25%), and covalently bound fatty acids (67-75%). Intragastric supplementation of EGF had no effect on the physicochemical changes caused by sialoadenectomy in the oral mucosal mucus coat, while nearly complete restoration to normal characteristics occurred in the gastric mucosal mucus coat. The results suggest that salivary EGF is essential for the maintenance of mucus coat dimension and quality needed in the protection of alimentary tract epithelium.

Animals↗
PubMed1988

Role of associated and covalently bound lipids in salivary mucin hydrophobicity: effect of proteolysis and disulfide bridge reduction.

The hydrophobic properties of salivary mucus glycoprotein were investigated by fluorescence spectroscopy using bis(8-anilino-1-naphthalene-sulfonate). The mucin, purified from rat submandibular salivary gland, was subjected to removal of associated and covalently bound lipids, degradation with pronase, and reduction with beta-mercaptoethanol, and titrated with the probe. Analyses of fluorescence data revealed the presence of 49 +/- 5 hydrophobic binding sites in the intact mucin molecule, a 69% increase in the number of binding sites occurred following extraction of associated lipids, while the removal of covalently bound fatty acids caused a 25% decrease in the binding sites. Proteolytic destruction of the nonglycosylated regions of the glycoprotein essentially abolished the probe binding, whereas reduction produced glycoprotein subunits whose combined number of hydrophobic binding sites was 2.4 times greater than that of mucus glycoprotein polymer. The results suggest that associated and covalently bound lipids contribute to hydrophobic characteristics of salivary mucin and that the hydrophobic binding sites reside on the nonglycosylated regions of this glycoprotein buried within its core.

Animals↗
PubMed1988

Enzymatic sulphation of mucus glycoprotein in rat sublingual salivary gland.

Sulphotransferase activity catalysing the transfer of sulphate ester group from 3'-phosphoadenosine-5'-phosphosulphate to salivary mucus glycoprotein was located in detergent extracts of the Golgi-rich membrane fraction of rat sublingual salivary glands. Optimum enzyme activity was obtained with 0.5 per cent Triton X-100, 20 mM NaF and 2 mM MgCl2, at pH 6.8, using desulphated sublingual salivary mucus glycoprotein. The enzyme was equally capable of sulphation of the proteolytically degraded and desulphated glycoprotein, whereas the acceptor capacity of intact salivary mucus glycoprotein was about four times lower. The Golgi enzyme preparation also catalysed the sulphation of galactosylceramide. However, the sulphation of mucus glycoprotein was not affected by the presence of this glycolipid, suggesting that the sulphotransferase involved in mucin sulphation is different from that responsible for the synthesis of galactosylceramide sulphate. The apparent Km of the sublingual-gland mucus glycoprotein sulphotransferase for salivary mucin was 7.7 microM. The 35S-labelled glycoprotein product of the enzyme reaction gave, in CsCl density gradient, a band in which the 35S label coincided with the glycoprotein. Alkaline borohydride reductive cleavage of this glycoprotein released the label into the reduced acidic oligosaccharide fraction. Upon thin-layer chromatography, two [35S]-oligosaccharides were detected. These were identified as penta- and heptasaccharides, each bearing a labelled sulphate ester group on the terminal N-acetylglucosamine residue. Based on the results of chemical and enzymatic analyses of the intact and desulphated compounds the following structures for these oligosaccharides are suggested: SO3----GlcNAc beta----Gal beta----GlcNAc beta----Gal beta----GlcNAc beta----(NeuAc----)GalNAc-ol and SO3----GlcNAc beta----Gal beta----GlcNAc beta----(NeuAc----)GalNAc-ol.

Animals↗
PubMed1988

Protection against alcohol-induced gastric mucosal injury by geranylgeranylacetone: effect of indomethacin.

The mechanism of gastric mucosal protection by an antiulcer agent, geranylgeranylacetone (GGA), against ethanol-induced injury was investigated. The experiments were conducted with groups of rats with and without intraperitoneal indomethacin pretreatment. Animals received intragastrically either a dose of GGA (200 mg/kg) or a vehicle, followed 30 min later by 1 ml of absolute ethanol. The rats were sacrificed after 30 min and the gastric mucosa was subjected to macroscopic and histologic assessment and the measurements of adherent mucus, its dimension and chemical composition. In the absence of GGA, ethanol produced advanced macroscopic necrosis (greater than 38%) and the extensive necrotic lesions were visible upon histologic examination. Pretreatment with GGA significantly reduced (p less than 0.001) the extent and depth of mucosal necrotic lesions caused by ethanol, and this protection was not thwarted by indomethacin. Evaluation of the adherent mucus and its dimension by Alcian blue uptake and inverted microscope technique revealed that GGA was also capable of preventing the untoward effect of indomethacin on the adherent gastric mucus gel and its thickness. Results of chemical analyses established that in the absence of GGA indomethacin caused an increase in mucus protein (15%) and a decrease in its covalently bound fatty acids (67%) and lipids (36%). The decrease in lipids was particularly reflected in the content of phospholipids. Indomethacin, however, had no apparent effect on the composition of gastric mucus elaborated in the presence of GGA. The results suggest that gastric mucosal protective action of GGA is not mediated by endogenous prostaglandins but rather appears to involve the metabolism of mucosal lipids.

Animals↗
PubMed1987

Enhancement of the lipid content and physical properties of gastric mucus by geranylgeranylacetone.

The effects of intragastric administration of geranylgeranylacetone (GGA) on the content, composition and physical properties of the mucus component of the gastric mucosal barrier were investigated. One group of rats received twice daily for 3 consecutive days a dose of 100 mg/kg body weight of GGA, while the control group was subjected to daily doses of the vehicle. Sixteen hours following the last dose, the animals were killed, and their stomach was cut open and subjected to measurements of the adherent mucus gel content, analysis of its lipids and molecular forms of elaborated mucin, and evaluation of the viscosity and H+ retardation capacity. The results revealed that GGA elicited a 62% increase in the adherent mucus gel and caused a marked decrease in the proportion of the lower molecular weight mucin. Furthermore, the mucus of the GGA group exhibited a 67% higher content of covalently bound fatty acids and contained 46% more total lipids which were greatly (143%) enriched in phospholipids. The physical measurements demonstrated that mucus elaborated in the presence of GGA also exhibited 2.3 times higher viscosity and had a 32% greater ability to retard the diffusion of H+ than the mucus of the control group. The results suggest that GGA exerts a profound effect on the lipid content and the properties of gastric mucus associated with the maintenance of the mucosal integrity.

Animals↗
PubMed1987

Campylobacter pyloridis degrades mucin and undermines gastric mucosal integrity.

The role of Campylobacter pyloridis, a spiral bacteria associated with gastritis and peptic ulcers in weakening the mucus component of gastric mucosal barrier was investigated. The colonies of bacteria, cultured from antral mucosal biopsies of patients undergoing gastroscopy, were washed with saline, passed through sterilization filter and the filtrate was examined for protease and glycosylhydrolase activities. The obtained results revealed that the filtrate exhibited a strong proteolytic activity not only towards the typical protein substrates such as albumin but also towards gastric mucin. Optimum enzymatic activity for degradation of mucin was attained at pH 7.0 and the protease activity was found in a low m.w. (less than 50K) protein fraction. The filtrate showed little glycosylhydrolase activity and did not cause the hydrolysis of mucin carbohydrates. The data suggest that C pyloridis infection weakens the gastric mucosal defense by causing proteolytic degradation of mucin component of the protective mucus layer.

Campylobacter↗
PubMed1986

Mucus glycoprotein of human saliva: differences in the associated and covalently bound lipids with caries.

A high molecular weight mucus glycoprotein has been isolated from submandibular saliva of caries-resistant and caries-susceptible individuals by a procedure involving fractionation on Bio-Gel P-100 and A-50 columns followed by equilibrium density-gradient centrifugation in CsCl. The purified caries-resistant mucus glycoprotein displayed a buoyant density of 1.50 and accounted for 9.5% of the dry weight of caries-resistant saliva. The caries-susceptible mucus glycoprotein represented 14.1% of the dry weight of caries-susceptible saliva and gave a buoyant density of 1.43. Both glycoproteins exhibited similar protein and carbohydrate content, but the caries-resistant mucus glycoprotein contained 28.7% less associated lipids and 3-times less covalently bound fatty acids than the caries-susceptible mucus glycoprotein. The associated lipids were represented by neutral lipids, glycolipids and phospholipids, whereas the covalently bound fatty acids consisted mainly of hexadecanoate, octadecanoate and docosanoate. Extraction of associated lipids caused the caries-resistant glycoprotein to band in CsCl gradient at the density of 1.54 and caused the caries-susceptible glycoprotein to band at the density of 1.52. A further shift in the buoyant densities occurred following removal of the covalently bound fatty acids, and both glycoproteins banded at the density of 1.57. While the intact caries-resistant and caries-susceptible glycoproteins were susceptible to proteolysis by pronase, the lipid-rich caries-susceptible glycoprotein was degraded to a lesser extent. Extraction of associated lipids increased the degradation of both glycoproteins, but the caries-susceptible glycoprotein still remained 25% less susceptible. However, the susceptibility to pronase of the delipidated and deacylated caries-resistant and caries-susceptible glycoproteins was essentially identical. The caries-resistant and caries-susceptible mucus glycoproteins also differed in susceptibility to peptic degradation. The apparent Km values for intact caries-resistant and caries-susceptible glycoproteins were 10.5 X 10(-7) M and 8.1 X 10(-7) M, while the values for the delipidated and deacylated caries-resistant and caries-susceptible glycoproteins were 13.0 X 10(-7) M and 12.4 X 10(-7) M. The results suggest that the differences in the content of associated lipids and covalently bound fatty acids are responsible for the different physiochemical characteristics of caries-resistant and caries-susceptible salivary mucus glycoproteins, which may be determining factors in the resistance to caries.

Chromatography, Gel↗
PubMed1986

In vitro fatty acid acylation of mucus glycoprotein from sublingual salivary glands.

The enzyme activity which catalyzes the transfer of palmitic acid from palmitoyl-coenzyme A to sublingual gland mucus glycoprotein has been demonstrated in the detergent extracts of the microsomal fraction of rat sublingual and parotid salivary glands. The acyltransferase activity of this fraction was similar in both types of glands. Further subcellular fractionation performed on sublingual glands revealed that the enzyme is associated with the Golgi-rich membrane fraction. Optimum enzymatic activity for fatty acylation of mucus glycoprotein was obtained using 0.5% Triton X-100, 2 mM dithiothreitol, 25 mM NaF, and 10 mM MgCl2 at a pH of 7.4. Higher concentrations of NaF, MgCl2 and dithiothreitol, however, were inhibitory. The apparent Km of the sublingual glands microsomal enzyme for mucus glycoprotein was 0.55 mg/ml and for palmitoyl-CoA, 3.5 X 10(-5) M. A 15% decrease in the acyltransferase activity was obtained with the reduced and alkylated mucus glycoprotein and it showed no activity towards the proteolytically degraded glycoprotein. The 14C-labeled product of the enzyme reaction gave in CsCl density gradient a band at the density of 1.49 in which the 14C label coincided with the glycoprotein. The 14C label in this glycoprotein was susceptible to deacylation with hydroxylamine, and the released labeled material was identified as palmitate.

Acylation↗