Biomedical subjects

A Slomiany

Publications and source records attributed to A Slomiany.

At least 127 records · Page 7Linked to original sources

PubMed1993

Pharmacological alterations of cellular transglutaminase activity and invasiveness in human colorectal carcinoma cells.

Human colorectal tumor cells expressing differing metastatic potential and tissue transglutaminase (TGA) activity were tested for the ability of various pharmacological agents to enhance TGA activity. The most effective stimulant was tetradecanoylphorbol-13-acetate (TPA), which in human colon carcinoma cells (SW620) caused a 5-fold, protein synthesis dependent increase in activity over 3 days. In WiDr and SW480 cells TGA activity was less susceptible to induction by TPA, possibly owing to the higher basal levels of TGA. Retinoic acid and a synthetic retinoid, [RO 15-1570; (E)-4-[2(5,6,7,8-tetramethylnaphthalene-2-yl)propen-1-yl] benzenesulphonyl-ethane)], also induced TGA activity to a lesser extent in SW620 cells, whereas other differentiation inducers [sodium butyrate and hexamethylene bis-acetamide (HMBA)] were ineffective. In LS174T cells, TGA activity was resistant to induction by all of the agents. The synthetic retinoid (RO 15-1570) inhibited in vitro invasiveness of SW620 cells, however, TPA treatment or addition of exogenous TGA did not inhibit invasiveness of these cells. Hence, the invasive behavior of a metastatic human colon tumor cell line (SW620) does not appear to be dependent on the TGA activity which the cells express. The anti-invasive activity of the retinoid in SW620 cells therefore may be mediated by some other mechanism.

Adenocarcinoma↗

PubMed1993

Effect of ebrotidine on gastric mucosal calcium channel activity.

Ebrotidine is a new H2-receptor antagonist also known for its gastroprotective effect against ethanol-induced mucosal injury. In this study, we investigated the effect of ebrotidine on the activity of the gastric mucosal calcium channels. The channel complex was isolated from the solubilized gastric epithelial cell membranes by affinity chromatography on wheat germ agglutinin. After being labeled with [3H]PN200-110, the complex was reconstituted into phosphatidylcholine vesicles which exhibited active 45Ca2+ uptake into intravesicular space and responded in a concentration-dependent manner to calcium channel activator, BAY K8644, as well as to calcium channel antagonist, PN200-110. The 45Ca2+ uptake was inhibited by ebrotidine. Maximum inhibitory effect was attained at 50 micrograms/ml ebrotidine, at which point a 54.9% decrease in uptake occurred. The gastric mucosal calcium channels, on epidermal growth factor binding (EGF) in the presence of ATP, responded by an increase in tyrosine phosphorylation of 55 and 170 kDa proteins, and the vesicles containing the phosphorylated channels displayed a 48% greater 45Ca2+ uptake. This phosphorylation process was inhibited by ebrotidine. Furthermore, ebrotidine also interfered with the binding of EGF to calcium channel protein. The results point toward the importance of EGF in the maintenance of gastric mucosal calcium homeostasis, and suggest that ebrotidine has the ability to protect the cellular integrity from calcium imbalance by modulating the EGF-stimulated gastric mucosal calcium channel phosphorylation.

Animals↗

PubMed1993

Effect of ebrotidine on gastric mucosal EGF and PDGF receptor expression.

The effect of ebrotidine, a new H2-blocker with gastroprotective properties, on the expression of gastric mucosal epidermal growth factor (EGF) and platelet-derived growth factor (PDGF) receptors, was investigated. Mucosal cell membranes were isolated from rats receiving twice daily for 5 days a dose of 100mg/kg ebrotidine or 100mg/kg ranitidine or vehicle only. Assays for EGF and PDGF revealed the presence of both types of receptors, activation of which led to enhanced tyrosine kinase activity. The receptor binding values in the control group were 2.4 fmol for EGF and 1.45 fmol/mg protein for PDGF, whereas the values in the ebrotidine group increased for EGF by 65.7% and 38.6% for PDGF, but no such effect was observed with ranitidine. The results suggest that the gastroprotective properties of ebrotidine stem from its ability to stimulate the epithelial proliferative activities through the enhancement of EGF and PDGF receptors expression.

Animals↗

PubMed1993

Identification of buccal mucosal mucin receptor.

A receptor for salivary mucin was isolated from epithelial cell membrane of buccal mucosa by a procedure involving membrane solubilization with octylglucoside followed by affinity chromatography on Sepharose-bound wheat germ agglutinin. The receptor protein displayed an apparent molecular weight of 97kDa and exhibited binding specific to mucin in a concentration-dependent manner. The receptor, furthermore, showed a requirement for carbohydrate chains in mucin for binding, as their removal caused a marked (89%) reduction in binding capacity. Scatchard analysis revealed a linear plot with a single class of high affinity binding (Kd = 1.1 microM; Bmax = 7.68nmol/mg protein. The results provide for the first time evidence for the existence in buccal mucosa of a specific receptor for salivary mucin.

Animals↗

PubMed1993

Enhancement in gastric mucosal laminin receptor expression with ulcer healing.

The expression of gastric mucosal laminin receptor with chronic ulcer healing was investigated. The receptor protein was isolated from gastric epithelial cell membrane of rats at various stages of ulcer healing and following radioiodination incorporated into vesicles which exhibited specific affinity towards laminin-coated surface. The binding assays revealed that the ulcer healing was accompanied by an increase in laminin receptor expression. A significant increase (2.5-fold) in the receptor expression occurred by the third day following the development of ulcer, reached a maximum of 8.6-fold increase by the 14th day when the ulcer was virtually healed, and its high level remained for at least 20 days. The results demonstrate the importance of laminin receptor as an indice of gastric mucosal repair in ulcer healing.

Acetates↗

PubMed1993

Characterization of gastric mucosal mucin receptor.

Mucin receptor was isolated from gastric epithelial cell membrane by a procedure involving membrane solubilization with octylglucoside followed by affinity chromatography on Sepharose-bound wheat germ agglutinin. The receptor protein yielded on SDS-PAGE a single 97kDa band and displayed specific affinity, in a concentration-dependent manner, towards the mucin-coated surface. The receptor showed requirement for carbohydrate chains in mucin for binding, as their removal caused a marked (87%) reduction in binding capacity. Scatchard analysis revealed a linear plot with a single class of high affinity binding (Kd = 43.8 nM; Bmax = 140 pmol/mg protein) sites. The results demonstrate for the first time the presence in gastric mucosa of a specific receptor for mucin.

Animals↗

PubMed1993

Mucus glycoprotein regulation of gastric mucosal calcium channel activity.

The effect of gastric mucin on the activity of calcium channel isolated from gastric epithelial cell membrane was investigated. The uptake of 45Ca2+ into the vesicle-reconstituted channels, while only moderately (14%) affected by the intact glycoprotein, was found significantly inhibited (59%) by the acidic glycoprotein fraction. This effect was associated with the sialic acid and sulfate ester groups of the glycoprotein. The channel complex in the presence of epidermal growth factor (EGF) and ATP responded by an increase in protein tyrosine phosphorylation, and the vesicles containing the phosphorylated channels showed a 50% increase in 45Ca2+ uptake. The channel protein phosphorylation was inhibited by the acidic mucus glycoprotein, which also interfered with the binding of EGF to the channel protein. The inhibitory effect was dependent upon the presence of sulfate ester and sialic acid groups, and the loss of the inhibitory capacity occurred following their removal. The results indicate that the acidic gastric mucins, by modulating the EGF-controlled calcium channel phosphorylation, play a major role in gastric mucosal calcium homeostasis.

Adenosine Triphosphate↗

PubMed1993

Salivary mucin modulation of buccal mucosal calcium channel activity.

The effect of the low and high molecular weight salivary mucins on the activity of calcium channel isolated from buccal epithelial cell membranes was investigated. The 45Ca2+ uptake into the vesicle-reconstituted channels, while only moderately (15%) affected by the intact mucin forms, was significantly inhibited (60%) by the acidic mucin fractions. This effect was associated with the sialic acid and sulfate ester groups of the glycoproteins. The channel complex in the presence of epidermal growth factor (EGF) and ATP responded by an increase in protein tyrosine phosphorylation, and the vesicles containing the phosphorylated channels showed a 46% increase in 45Ca2+ uptake. The phosphorylation process was inhibited by the acidic mucins, which also interfered with the binding of EGF to the channel protein. The inhibitory effect was dependent upon the presence of sulfate ester and sialic acid groups, and the loss of the inhibitory capacity occurred following their removal. The results demonstrate the salivary mucins actively participate in the modulation of buccal mucosal calcium channel activity.

Adenosine Triphosphate↗

PubMed1993

Glycosulfatase activity of Porphyromonas gingivalis a bacterium associated with periodontal disease.

Porphyromonas gingivalis, a bacterium implicated in the pathogenesis of periodontal disease, was found to elaborate an extracellular glycosulfatase enzyme. Upon purification by low temperature acetone fractionation, an active enzyme at 60% acetone was obtained which on SDS-PAGE gave a protein band of 37kDa. The glycosulfatase effectively caused desulfation of galactosyl- and lactosylceramide sulfates (pH 5.0) which contain the sulfate ester groups at C-3 of galactose, a well as proteoglycans (pH 5.7-6.2) of gingival tissue which are rich in N-acetylgalactosamine-4-sulfate, but not the sulfated salivary mucin with the sulfate groups at C-6 of galactose and C-6 of N-acetylglucosamine. The results demonstrate for the first time that P. gingivalis displays glycosulfatase activity and that the disruptive action of this enzyme may be a major factor in the etiology of periodontal disease.

Culture Media↗

PubMed1993

Salivary and plasma insulin levels in man.

Changes in plasma and salivary immunoreactive insulin were studied in saliva and blood of 9 healthy subjects following either oral glucose or a test meal ingestion at 30 min intervals for up to 3h. The levels of plasma immunoreactive insulin reached maximum at 60 min with a value of 62.77 +/- 6.52 microU/ml after the glucose load and 62.62 +/- 6.52 microU/ml after the test meal. In the case of salivary immunoreactive insulin levels, the maximum was attained at 90 min with a value of 18.47 +/- 2.68 microU/min after the glucose load and 22.40 +/- 2.01 microU/min after the test meal. Positive linear correlation (r = 0.74) was found between the salivary and plasma immunoreactive insulin levels.

Administration, Oral↗

PubMed1993

Inhibition of Helicobacter pylori colonization by an antiulcer agent, sulglycotide.

Sulglycotide, a potent antiulcer agent derived from duodenal mucus glycopeptide through sulfation of the carbohydrate moieties, was evaluated with respect to its ability to interfere with H. pylori mucosal attachment. H. pylori cells were incubated with sulglycotide or human gastric mucin and then examined for their inhibitory capacity of H. pylori attachment to erythrocytes. Titration data revealed that the mucin inhibitory activity was confined to its sulfomucin fraction, the titer of which was found to be 16-fold higher than that of intact mucin. The data with sulglycotide showed that the inhibitory titer of this agent against H. pylori attachment was at least 30-fold higher than that of the sulfated gastric mucin fraction. The results point towards the involvement of sulfomucins in the protection of gastric mucosa from H. pylori colonization and demonstrate that sulglycotide, because of structural similarities, is ideally suited to augment the inherent mucosal defenses against this pathogen.

Anti-Ulcer Agents↗

PubMed1993

Glycosulfatase activity of Helicobacter pylori towards gastric sulfomucin: effect of nitecapone.

A glycosulfatase activity towards human gastric sulfomucin was identified in the extracellular material elaborated by H. pylori, a pathogen implicated in the etiology of gastric disease. The purified enzyme exhibited maximum activity at pH 5.7 in the presence of 0.3% Triton X-100 and 100mM CaCl2, and displayed on SDS-PAGE an apparent molecular weight of 30kDa. The H. pylori glycosulfatase effectively caused desulfation of N-acetylglucosamine-6-sulfate and galactose-6-sulfate of the carbohydrate chains of mucins, as well as that of glucose-6-sulfate of glyceroglucolipids, but was ineffective towards galactosyl -and lactosylceramide sulfates which contain galactose-3-sulfate. The glycosulfatase activity towards human gastric sulfomucin was inhibited by an antiulcer agent, nitecapone, which at its optimal concentration (100 micrograms/ml) caused a 61% decrease in mucin desulfation. The results show that H. pylori through its glycosulfatase activity causes desulfation of sulfated mucins and glyceroglucolipids of the protective mucus layer, and that nitecapone is able to interfere with this detrimental action.

Anti-Ulcer Agents↗

PubMed1993

Effects of exogenous transglutaminase on spreading of human colorectal carcinoma cells.

Using pre-confluent cultures of a human colon tumor cell line deficient in transglutaminase (LS174T cells), we have investigated the effect of adding exogenous transglutaminase (TGA) on cell spreading. The cells were plated at either 4.5 x 10(5) cells per well (low-seeded cultures) or 9 x 10(5) cells per well (high-seeded cultures) in 24-well dishes and treated for either 1 or 4 days (low- and high-seeded cultures respectively) under following conditions: Chee's Essential Medium (CEM) + 10% fetal calf serum (FCS); CEM + 10% FCS + TGA; CEM + 10% FCS + dithiothreitol (DTT) + CaCl2; CEM + 10% FCS + DTT + CaCl2 + TGA. Photomicrography of the cells after these treatments revealed that in both low- and high-seeded cultures, TGA inhibited the spreading of the cells both in the presence and absence of DTT and calcium. Individual colony sizes were significantly smaller in the presence of TGA. This phenomenon may be related to the ability of TGA to promote cell interactions with the underlying tissue matrices and metastasis.

Calcium Chloride↗

PubMed1992

Effect of cigarette smoking on salivary epidermal growth factor (EGF) and EGF receptor in human buccal mucosa.

The mouth acts as a primary target for cigarette smoke which is associated with several oral diseases and cancer. The present study investigated the effect of cigarette smoking on salivary EGF and the buccal EGF receptor. Samples of whole saliva and buccal biopsy were obtained from 15 healthy volunteers (10 smokers and 5 non-smokers). The smokers smoked 20 or more cigarettes/day for more than 5 years. Salivary cotinine (a major metabolite of nicotine) was determined by radioimmunoassay (RIA). The salivary cotinine level was consistent with the self-reported smoking status (smokers, 106-530 ng/ml saliva; non-smokers, < 2 ng/ml saliva). As compared to the non-smokers, the salivary EGF concentration (determined by RIA) was 32% lower in those smokers whose salivary cotinine level was 250 ng/ml or higher (non-smokers, 2.21 +/- 0.16; smokers, 1.57 +/- 0.09 ng/ml saliva; mean +/- S.E.M., P < 0.01). There was no significant difference in 125I-labeled EGF binding to the buccal receptor between the two groups. However, EGF stimulated the autophosphorylation of a 170-kDa protein band in the sample of non-smokers, but not in the smokers. The immunoblot analysis using anti-EGF receptor antibody indicated that the smoking-related deficiency in EGF receptor autophosphorylation was due to the functional alteration of the receptor proteins. In conclusion, cigarette smoking reduces the salivary EGF level and impairs the function of buccal EGF receptor, which may be associated with the pathology of smoking-related oral disease.

Blotting, Western↗

PubMed1992

Platelet-derived growth factor activation of gastric mucosal calcium channels.

Gastric mucosal calcium channel complex was isolated from the solubilized epithelial cell membranes by affinity chromatography on wheat germ agglutinin. The complex following labeling with [3H]PN200-100 was reconstituted into phospholipid vesicles which exhibited active 45Ca2+ uptake. The channels responded in a dose dependent manner to dihydropyridine calcium antagonist, PN200-110, which at 0.5 microM exerted maximal inhibitory affect of 66% on 45Ca2+ uptake, while a 52% enhancement in 45Ca2+ uptake occurred with a specific calcium channel activator, BAY K8644. On platelet-derived growth factor (PDGF) binding in the presence of ATP, channels showed an increase in protein tyrosine phosphorylation of 55 and 170kDa subunits of calcium channel. Such phosphorylated channels following reconstitution into vesicles displayed a 78% greater 45Ca2+ uptake. The results point towards the importance of PDGF in the regulation of gastric mucosal calcium homeostasis.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

PubMed1992

Helicobacter pylori lipopolysaccharide effect on the synthesis and secretion of gastric sulfomucin.

The effect of H. pylori lipopolysaccharide on the synthesis and secretion of sulfated mucin in gastric mucosa was investigated using mucosal segments incubated in the presence of [3H]proline, [3H]glucosamine and [35S]Na2SO4. The lipopolysaccharide, while showing no discernible effect on the apomucin synthesis was found to inhibit the process of mucin glycosylation and sulfation, which at 100 micrograms/ml lipopolysaccharide reached the optimal inhibition of 65%. The analysis of mucin secretory responses revealed that the lipopolysaccharide by first 15 min caused a 57% stimulation in sulfomucin secretion followed thereafter by inhibition, which reached maximum of 32% by 45 min. The results suggest that colonization of gastric mucosa by H. pylori may be detrimental to the process of gastric sulfomucin synthesis and secretion.

Animals↗

PubMed1992

Glycosulfatase activity of Helicobacter pylori toward gastric mucin.

A glycosulfatase activity toward sulfated gastric mucus glycoprotein was identified in the extracellular material elaborated by H. pylori, a bacteria implicated in the etiology of gastric disease. Upon acetone precipitation, an active enzyme fraction at 64% acetone was obtained which on SDS-PAGE gave a major 30kDa protein band. The H. pylori glycosulfatase exhibited maximum activity (314.8 pmol/mg protein/h) at pH 5.7 in the presence of Triton X-100 and CaCl2, and was capable of removal of the sulfate ester groups situated at C-6 of N-acetylglucosamine, galactose and glucose. However, the enzyme was ineffective toward galactosylceramide and lactosylceramide sulfates which contain the sulfate ester group on C-3 of galactose. The results suggest that H. pylori is capable of overcoming the interference by sulfated mucus glycoprotein with its colonization of gastric mucosa.

Gastric Mucins↗

PubMed1992

Role of adrenergic and cholinergic mediators in salivary phospholipids secretion.

The influence of adrenergic and cholinergic mediators on phospholipid secretion in rat sublingual salivary gland cells maintained in the presence of [3H]choline was investigated. The secretion of [3H]choline-containing phospholipids over 30 min period averaged 1.93% of the total cellular labeled phospholipids in the absence of any mediator, and was enhanced by beta-adrenergic agonist, isoproterenol, to a greater extent than the cholinergic agonists, pilocarpine and carbachol. A 2.9-fold increase in phospholipid secretion occurred with isoproterenol, while pilocarpine and carbachol evoked only 1.3-fold increase. The effect of isoproterenol was inhibited by alprenolol and that of pilocarpine and carbachol by atropine. In contrast to pilocarpine and carbachol, the enhanced phospholipid secretion due to isoproterenol was accompanied by an increase in cAMP concentration. The secretion of phospholipids was also stimulated by dibutyryl-cAMP and the protein kinase C activator, phorbol myristate acetate, but not by 4 alpha-phorbol 12,13-didecanoate which does not activate protein kinase C. Furthermore, the effects of dibutyryl-cAMP and phorbol myristate acetate were additive. The phospholipids secreted in response to isoproterenol exhibited a 52% decrease in lysophosphatidylcholine, while those secreted in response to pilocarpine and carbachol showed a 21-23% lower content of phosphatidylcholine, and were enriched in lysophosphatidylcholine (2.6-2.8-fold) and sphingomyelin (1.5-1.6-fold). The results indicate that salivary phospholipid secretion remains mainly under beta-adrenergic regulation, while the phospholipid makeup of the secretion is under cholinergic control.

Adrenergic beta-Agonists↗