sodium-dodecyl-sulfate and Mouth-Neoplasms

Toothpastes contain three main components: detergents, abrasives, and fluoride. Detergents, particularly sodium lauryl sulfate, have been proposed as components that enable toothpastes to produce cytotoxic effects in vitro. However, not all toothpastes contain sodium lauryl sulfate, and almost no studies have found an association between detergents and the in vitro cytotoxicity of toothpastes. The present study examined the in vitro cytotoxicity of nine commercially available toothpastes containing four different detergents. Toothpastes were diluted in serum-free medium, centrifuged, and filter sterilized. The half-lethal concentration of the toothpaste-conditioned medium (TCM) was calculated based on the formation of formazan by gingival fibroblasts, oral squamous cell carcinoma HSC-2 cells, and L929 cells. Cell proliferation was analyzed, and live-dead staining was performed, after exposure of cells to conditioned medium prepared with 1% toothpaste (1% TCM). It was found that toothpastes containing sodium lauryl sulfate and amine fluoride strongly inhibited cell viability with the half-lethal concentration being obtained with conditioned medium prepared with approximately 1% toothpaste (1% TCM). Toothpastes containing cocamidopropyl betaine and Steareth-20 showed higher half-lethal concentration values, with the half-lethal concentration being obtained with conditioned medium prepared with 10% (10% TCM) and 70% (70% TCM) toothpaste, respectively. Proliferation and live-dead data were consistent with the cell-viability analyses. These results demonstrate that the type of detergent in toothpastes can be associated with changes in in vitro cell toxicity.

Topics: Animals; Betaine; Carcinoma, Squamous Cell; Cariostatic Agents; Cell Culture Techniques; Cell Line; Cell Line, Tumor; Cell Proliferation; Cell Survival; Cells, Cultured; Culture Media, Conditioned; Culture Media, Serum-Free; Detergents; Diamines; Epithelial Cells; Fibroblasts; Fluorescent Dyes; Fluorides; Formazans; Gingiva; Humans; Indicators and Reagents; Materials Testing; Mice; Mouth Neoplasms; Polyethylene Glycols; Sodium Dodecyl Sulfate; Tetrazolium Salts; Toothpastes

The effect of sodium lauryl sulphate (SLS) on cytokeratin (CK) gene expression in hamster cheek pouch epithelium was studied with a hybridohistochemical technique. Using specific human anti-sense RNA probes, the plausible hamster mRNA counterparts for these human CK mRNAs were localized by detection of heterologous hybrids. In comparison with normal epithelium, the expression and distribution pattern of CK mRNAs in the hamster cheek pouch were obviously changed after application of SLS. There was a decreased expression of CK mRNAs in the hyperplastic basal layer, and increased expression in the hypertrophic granular layer. Strikingly, hybridization with the human CK 18 cRNA probe revealed an additionally expressed CK mRNA in the SLS-treated epithelium that was not found in the untreated epithelium. The present study indicates that cRNA probes for human CK mRNAs can be used successfully, not only to distinguish between different hamster CK mRNAs but also to investigate changes in CK gene expression upon the induction of non-neoplastic and neoplastic alterations in the hamster cheek pouch model. This may help elucidate the molecular changes involved in epithelial pathologies.

Topics: Animals; Cheek; Cricetinae; Detergents; Epithelium; Gene Expression Regulation; Gene Expression Regulation, Neoplastic; Histocytochemistry; Humans; Hybridomas; Hyperplasia; Hypertrophy; In Situ Hybridization; Keratins; Male; Mesocricetus; Mouth Mucosa; Mouth Neoplasms; RNA Probes; RNA, Antisense; RNA, Complementary; RNA, Messenger; Sodium Dodecyl Sulfate; Surface-Active Agents

Urokinase-type (uPA) and tissue-type (tPA) plasminogen activators were identified by fibrinolytic autography in the sulcus epithelium of human gingival mucosa but not in the orthokeratinized gingival epithelium. Fibrinolytic activity was present only over blood vessels in frozen sections of oral squamous cell carcinomas, the malignant epithelial cells showing no plasminogen activator activity. Plasminogen activators could not be demonstrated in either the sulcus or gingival epithelium by immunofluorescence, but both uPA and tPA were found in occasional squamous carcinoma cells. Fibrinolytic activity of culture fluids from epithelial explants grown in vitro from human gingival mucosa showed marked variation, but activity was much higher in the culture supernatants than in the cell lysates. Fibrinolytic activity of culture fluids from epithelial explants of squamous cell carcinomas was low both in supernatants and lysates. Zymogram overlays of sodium dodecyl sulphate-polyacrylamide electrophoretic gels from culture supernatants showed that the low fibrinolytic activity of culture supernatants of oral squamous cell carcinomas was due to the associated presence of plasminogen activator inhibitors. The fibrinolytic activity in the zymogram was due predominantly to uPA but some lysis was due also to tPA.

Topics: Carcinoma, Squamous Cell; Cells, Cultured; Connective Tissue; Electrophoresis, Polyacrylamide Gel; Epithelial Cells; Epithelium; Fibrinolysis; Fluorescent Antibody Technique; Gingiva; Humans; In Vitro Techniques; Keratinocytes; Molecular Weight; Mouth Neoplasms; Sodium Dodecyl Sulfate; Tissue Plasminogen Activator; Urokinase-Type Plasminogen Activator

Cytokeratin pattern was analyzed in 14 moderately differentiated and 12 well-differentiated squamous cell carcinomas of buccal mucosa by SDS-PAGE, immunoblotting and two dimensional electrophoresis. These were compared with patterns of normal buccal mucosa and surrounding areas whenever possible. Normal buccal mucosa expresses keratin No. 4 (59Kd), 5 (58Kd), 13 (54Kd) and 14 (50Kd). Keratin No. 4 (59Kd) and 14 (50Kd) were expressed by 20 of 26 tumors studied, while many of the tumors did not express keratins No. 5 (58Kd) and 13 (54Kd). Keratin No. 1 (67Kd) and 16 (48Kd) were aberrantly expressed by 9 well-differentiated tumors. Keratin No. 17 (46Kd) and 18 (45Kd) were expressed by 10 and 8 tumors of 14 moderately differentiated tumors. Six tumors which showed involvement of alveolar mucosa, expressed some keratins expressed by its normal counterpart. Their altered expression was consistent with the differentiation pattern as stated earlier. Non-expression of keratins 5 and 13 seems to be the result of malignant transformation and is seen in the majority of tumors, while appearance of aberrant keratins seems to be related more to the degree of differentiation of the tumor.

Topics: Carcinoma, Squamous Cell; Electrophoresis, Gel, Two-Dimensional; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Humans; Immunoblotting; Isoelectric Focusing; Keratins; Mouth Mucosa; Mouth Neoplasms; Sodium Dodecyl Sulfate

The pattern of keratin expression in hamster cheek pouch epithelium during 15-wk of DMBA-induced carcinogenesis was studied. The sequential changes in cytokeratins of premalignant and malignant tissues and comparative investigation of normal epithelial tissues were examined during a weekly sequential DMBA-induced chemical carcinogenesis. Keratin polypeptides of normal pouch epithelium appear in a molecular weight range of 43-67 kd and 5-6 proteins can be identified. The disappearance of high molecular weight keratin (61-67 kd) was observed from the 6-wk DMBA-treated premalignant group to the 15-wk DMBA-treated malignant group. An additional keratin polypeptide was noted initially on the 11th-wk-DMBA-treated group and remained to the 15th-wk-DMBA treated group.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Cheek; Cricetinae; Electrophoresis, Polyacrylamide Gel; Epithelium; Immunoblotting; Keratins; Male; Mesocricetus; Mouth Mucosa; Mouth Neoplasms; Palate; Precancerous Conditions; Skin; Sodium Dodecyl Sulfate

The frequency of the occurrence of cytokeratins analyzed SDS-electrophoretically in 20 leukoplakic lesions and 14 squamous cell carcinomas of the oral mucosa has shown a picture of "restlessness" with some quantitative, some qualitative deviations from the locally normal pattern of cytokeratins. In most cases the basic pattern typical for the oral mucosa was still recognizable, except for three highly undifferentiated carcinomas. The variations of the cytokeratin pattern existed independently of the clinical form of leukoplakia and of its histological degree of dysplasia. The striking findings in some, but not all cases were: --the presence of cytokeratin no. 18 as a major component which normally appears rarely and faintly, and of cytokeratin no. 19, which is not normally detectable electrophoretically within the whole oral mucosa; --the absence of cytokeratins no. 1-3 despite of the cornification which was reliably proved by histology; --the appearance of the proteins having molecular mass values of 42 kDa and less which very probably may be the products of partial keratinolysis as evidenced by immunoblotting with monoclonal and polyclonal antibodies to cytokeratins. Among these proteins a proteolytically modified cytokeratin no. 19 of only 38 kDa was found.

Topics: Antibodies, Monoclonal; Biomarkers, Tumor; Carcinoma, Squamous Cell; Electrophoresis, Polyacrylamide Gel; Humans; Immunoblotting; Keratins; Leukoplakia, Oral; Mouth Mucosa; Mouth Neoplasms; Precancerous Conditions; Sodium Dodecyl Sulfate

Treatment with sodium dodecyl sulfate (SDS) converted the vaccinia virus strain IHD-J into particles of two types: (i) ghosts which possessed a thin-membrane vesicle derived from basement part of the virus membrane with attached lateral bodies and a membranous structure derived from the core wall and (ii) aggregates of a DNA-nucleoprotein eluted from the core. These particles lacked lipids, and all the viral phospholipids were detected in the SDS-soluble fraction. The viral membrane was composed of an SDS-soluble coat layer and the basement membrane, and the basement membrane was maintained by a mechanism other than the lipid bilayer. By comparisons of protein species in morphologically distinct subviral particles prepared by several solubilizing methods, protein compositions of viral structural elements were suggested as follows: 25,000-molecular-weight viral protein-17,000-molecular-weight viral protein ( VP25K - VP17K ), viral basement membrane; VP13 . 8K , major component of the lateral body; VP70K , VP69K , VP66K , and VP64K , minor components of the lateral body; VP61K , outer layer of core wall; VP57K - VP22K , inner layer of core wall; and VP27K - VP13K , nucleoprotein. These structural elements found in the SDS-insoluble particles dissolved in the same SDS solution under reducing conditions, indicating that the disulfide linkages seem to have a principal role in maintaining their morphological integrity. VP57K , VP27K , VP13 . 8K , and VP13K were revealed to possess affinity for DNA. Denatured calf thymus DNA and viral DNA in double- or single-stranded form associated equally well with these proteins, but RNA did not bind. Therefore, it was strongly suggested that disulfide-linked VP27K - VP13K represented the nucleoproteins of vaccinia virus. A structural model of vaccinia virus is proposed and discussed.

Topics: Carcinoma; Cell Line; Disulfides; DNA-Binding Proteins; Electrophoresis, Polyacrylamide Gel; Humans; Microscopy, Electron; Molecular Weight; Mouth Neoplasms; Sodium Dodecyl Sulfate; Trypsin; Vaccinia virus; Viral Proteins

Topics: Adenoviridae; Carcinoma; Cell Line; Centrifugation, Density Gradient; Chromatography, Affinity; Edetic Acid; Electrophoresis, Polyacrylamide Gel; Formamides; Humans; Molecular Weight; Mouth Neoplasms; Nucleoproteins; Peptides; Polyribosomes; RNA, Messenger; RNA, Ribosomal; Sodium Dodecyl Sulfate; Sulfur Radioisotopes; Tritium

Topics: Adenoviridae; Anti-Bacterial Agents; Autoradiography; Carcinoma; Cell Line; Cytarabine; Electrophoresis, Polyacrylamide Gel; Guanidines; HeLa Cells; Methionine; Mouth Neoplasms; Nucleosides; Peptide Biosynthesis; Poliovirus; Sodium Dodecyl Sulfate; Sulfur Radioisotopes; Time Factors; Virus Replication

Topics: Adenoviridae; Animals; Carcinoma; Carcinoma, Krebs 2; Cell Line; Cell-Free System; Centrifugation, Density Gradient; Chromatography, DEAE-Cellulose; Chromatography, Ion Exchange; Electrophoresis; Electrophoresis, Polyacrylamide Gel; Hydroxyapatites; Magnesium; Methionine; Mice; Mouth Neoplasms; Peptide Fragments; Phosphates; Polyribosomes; Protein Biosynthesis; Sodium Dodecyl Sulfate; Spectrophotometry, Ultraviolet; Sulfur Radioisotopes; Tritium; Trypsin; Uridine; Viral Proteins

Topics: Adenoviridae; Carcinoma; Cell Line; Defective Viruses; Electrophoresis, Polyacrylamide Gel; Mouth Neoplasms; Peptides; Sodium Dodecyl Sulfate; Thymidine; Time Factors; Tritium; Virus Replication

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified incomplete particles of adenoviruses type 2 and 3 revealed that core proteins V and VII and capsid proteins VI, VIII, and X were absent in these particles, but they contained polypeptides not present in complete particles. Two types of incomplete particles were observed in the electron microscope, appearing as deoxyribonucleic acid-less particles with single discontinuities in the capsid structure (about 80%), or more amorphous particles resembling hexon aggregates (about 20%). The amount of incomplete and complete particles increased in parallel during the infectious cycle. Detectable amounts were found at 13 h with a maximum rate of synthesis for both particles at 24 h after infection. (3)H-labeled amino acids were incorporated into incomplete particles without a detectable lag period, but the label appeared in complete particles with a 60- to 80-min lag. Early after the pulse in pulse-chase experiments, the radioactivity was higher for incomplete particles than for complete particles and leveled off before the activity of complete particles reached a maximum. In the adenovirus type 2 system, pulse-chase experiments suggested a precursor-product relationship between incomplete and complete particles. After a short pulse, 19 h postinfection, entrance of (3)H-labeled amino acids into the hexon polypeptide of complete particles was delayed for 80 min, but no delay was observed for the labeling of the hexon polypeptide of incomplete particles. The core polypeptides appear in complete particles without a delay, also suggesting that incomplete particles were precursors to complete particles. Incorporation of (3)H-labeled amino acids into the hexon polypeptide of complete and incomplete particles was drastically decreased by inhibition of protein synthesis with emetine. However, the uptake of label into core proteins of complete particles was only decreased to 50% on inhibition of protein synthesis. The results suggest that incomplete particles are intermediates in virus assembly in vivo and that the assembly of capsid polypeptides into incomplete and complete particles is dependent on continuing protein synthesis.

Topics: Adenoviridae; Amino Acids; Carcinoma; Centrifugation, Density Gradient; Electrophoresis, Polyacrylamide Gel; HeLa Cells; Humans; Inclusion Bodies, Viral; Microscopy, Electron; Mouth Neoplasms; Peptides; Protein Precursors; Sodium Dodecyl Sulfate; Time Factors; Tritium; Viral Proteins; Virus Cultivation; Virus Replication; Viruses

Topics: Adenoviridae; Animals; Arginine; Autoradiography; Carcinoma; Cell Fractionation; Cell Line; Cells, Cultured; Cytarabine; Electrophoresis, Disc; Humans; Immunodiffusion; Kidney; Methionine; Mouth Neoplasms; Peptide Biosynthesis; Peptides; Rabbits; Sodium Dodecyl Sulfate; Sulfur Isotopes; Time Factors; Viral Proteins

Topics: Amino Acids; Animals; Bacteriophages; Carbon Isotopes; Carcinoma; Cattle; Cell Line; Centrifugation, Density Gradient; Coliphages; Corynebacterium diphtheriae; Diphtheria Toxin; Electrophoresis, Disc; Humans; Iron; Mouth Neoplasms; Neoplasm Proteins; Serum Albumin, Bovine; Sodium Dodecyl Sulfate; Trypsin; Viral Proteins; Virus Cultivation

Three major structural proteins were found in adenovirus-associated virus (AAV) type 3H virions which were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The molecular weights of the polypeptides were determined to be approximately 66,000 (VP1), 80,000 (VP2), and 92,000 (VP3). The component having a molecular weight of 66,000 comprised about 80% of the total virion protein in the major AAV-3H particle, and the other two components comprised about 10% each. Proteins of the same molecular weight were found in the minor dense AAV-3H virion, but the 80,000- and 92,000-molecular-weight components were present at about one-half the concentration. The AAV-3H virion contains about 72 molecules of VP1 and 8 and 7 molecules of VP2 and VP3, respectively.

Topics: Adenoviridae; Carcinoma; Cell Line; Dithiothreitol; Electrophoresis, Polyacrylamide Gel; Humans; Molecular Weight; Mouth Neoplasms; Peptides; Satellite Viruses; Serotyping; Sodium Dodecyl Sulfate; Viral Proteins