phosphorus-radioisotopes and Mouth-Neoplasms

Combined modality therapy plays a central role in the management of advanced head and neck tumors. The objective of our Phase II study was to determine the feasibility, toxicity, and clinical and pathologic response of preoperative induction chemotherapy, followed by concurrent chemoradiotherapy in patients with Stage III or IV squamous cell carcinoma according to the American Joint Committee on Cancer Staging of the oral cavity and oropharynx with no distant metastases.. After staging, 62 patients with locally advanced carcinoma of the oral cavity and oropharynx were treated preoperatively with chemotherapy (1 cycle of cisplatin and 5-fluorouracil [P-5FU]) followed by concurrent chemoradiotherapy (3 cycles of P-5FU combined with radiotherapy, 60 grays [Gy] given in 33 fractions of 1.8 Gy). After evaluation, patients underwent surgery either as a diagnostic (biopsy) or therapeutic procedure (resection of the primary tumor and/or the neck). Surgery was performed with the intent to spare organ function.. Grade 3-4 mucositis was observed in 37 patients (59%). Overall clinical response was obtained in 87%, and the complete clinical response rate was 50%. Surgery was performed in 53 patients, 50 at the primary tumor site (11 biopsies, 14 marginal excisions, and 25 wide excision) and 46 patients had neck dissection. Pathologic complete remission was observed in 29 patients (46%). After a median follow-up of 39 months, locoregional control rate was 76%, estimated 3-year disease free survival rate was 73% (+/- 4%), and estimated 3-year overall survival rate was 76% (+/- 4%).. This intensive multimodality treatment is feasible, and toxicity is significant but tolerable. The treatment results appear promising and durable. Organ-preserving surgery can be performed in many patients.

Topics: Adult; Aged; Antineoplastic Agents; Cisplatin; Combined Modality Therapy; Feasibility Studies; Female; Fluorouracil; Humans; Male; Middle Aged; Mouth Neoplasms; Oropharyngeal Neoplasms; Phosphorus Radioisotopes; Preoperative Care; Radiography; Radiopharmaceuticals; Survival Analysis; Treatment Outcome

Samples of clinically normal oral tissue were obtained from 17 tobacco smokers and 7 non- or ex-smokers undergoing surgery for intra-oral squamous cell carcinoma. Isolated DNA was analysed for the presence of aromatic DNA adducts using the 32P-postlabelling technique with adduct enhancement by either butanol extraction or nuclease P1 enrichment. DNA adduction detected following butanol extraction was more diverse and at a higher level than obtained with the P1 method. Adduct levels in the smokers and non- or ex-smokers were 1163 +/- 375 and 774 +/- 318 amol/micrograms, respectively. This difference is statistically significant (p < 0.05). The differential enhancement of adducts with the two protocols suggested that arylamines may be the source of at least a proportion of the DNA adduction detected. These data indicate that oral tissues are likely to be a suitable source of material for human biomonitoring and furthermore they highlight the importance of utilizing more than one enhancement procedure when examining DNA adduction induced by complex mixtures such as tobacco smoke or those encountered at industrial plants.

Topics: Butanols; Carcinoma, Squamous Cell; DNA; DNA Damage; DNA, Neoplasm; Environmental Monitoring; Evaluation Studies as Topic; Humans; Mouth Mucosa; Mouth Neoplasms; Phosphorus Radioisotopes; Single-Strand Specific DNA and RNA Endonucleases; Smoking

Samples of clinically normal oral tissue were obtained from patients undergoing surgery for intraoral squamous cell carcinoma. DNA was extracted from samples obtained from 20 tobacco smokers, four exsmokers, and nine nonsmokers and analyzed for the presence of aromatic DNA adducts using two distinct modifications of the 32P postlabeling assay. 32P postlabeling following butanol extraction enhancement revealed a much wider range and substantially higher levels of DNA adducts than obtained following nuclease P1 enrichment. Adduct levels in smokers, exsmokers, and nonsmokers were 1133 +/- 354, 785 +/- 251, and 660 +/- 317 amol/microgram of DNA (+/- SD), respectively. The elevation of adduct levels in smokers compared with either nonsmokers or non- and exsmokers combined is statistically significant (P < 0.005). These observations are consistent with epidemiological evidence linking tobacco smoking with oral cancer. The differential enhancement of DNA adducts with the two 32P postlabeling protocols indicate that aromatic amines and nitroaromatics may be important sources of the DNA adducts detected in human oral tissue.

Topics: Adolescent; Adult; Age Factors; Aged; Aged, 80 and over; Alcohol Drinking; Butanols; Carcinoma, Squamous Cell; DNA; DNA Damage; Female; Humans; Male; Middle Aged; Mouth; Mouth Neoplasms; Phosphorus Radioisotopes; Single-Strand Specific DNA and RNA Endonucleases; Smoking

Formation of carcinogen-DNA adducts in rat oral epithelial cells after treatment with cigarette smoke condensate (CSC) or chewing tobacco in the presence of ethanol was investigated using the 32P-postlabeling procedure. Concomitant treatment of the cells with ethanol increased the relative adduct level over that found in cells treated with tobacco smoke condensate only. Treatment with chewing tobacco resulted in slightly higher adduct levels than in controls. Treatment of the cells with ethanol did not significantly increase the uptake of a polycyclic aromatic hydrocarbon, benzo[j]fluoranthene, however, high tar CSC alone or in combination with ethanol significantly increased the uptake of radiolabeled benzo[j]fluoranthene, suggesting that increased uptake of the carcinogens may be one of the synergistic mechanisms of alcohol in oral carcinogenesis.

Topics: Animals; Carcinogens; Cheek; Cocarcinogenesis; DNA; DNA Damage; Drug Synergism; Ethanol; Fluorenes; Isotope Labeling; Mouth Mucosa; Mouth Neoplasms; Phosphorus Radioisotopes; Plants, Toxic; Rats; Smoke; Tobacco, Smokeless

Development of oral cavity cancer in man has been linked to alcohol consumption and use of tobacco products. In order to understand the underlying carcinogenic mechanisms in the oral cavity a method is needed to monitor exposure of this site to various environmental insults. In this pilot study we evaluate the use of the 32P-postlabeling assay to detect adducts in DNA from exfoliated oral mucosa cells. Exfoliated cells were collected from the cheek and tongue of 27 men aged 35-69 years. DNA was extracted from the cells and analyzed by the enhanced 32P-postlabeling technique using butanol extraction. A variety of adduct spots were detected but none was consistently associated with exposure to alcohol or tobacco products. Some of the adducts detected had migration patterns in TLC very similar to the major deoxyguanosine adducts formed by the diol epoxides of benzo[a]pyrene and 5-methylchrysene, suggesting that they may have been formed from polynuclear aromatic hydrocarbons. Adduct spots with migration patterns similar to polynuclear hydrocarbon adducts accounted for only about one third of the total adduct spots observed. Relative adduct labeling (RAL) values were determined for samples from 12 of the 27 individuals. RAL values ranged from 1.6 X 10(-6) to 7.7 X 10(-11) adducts per nucleotide. The RAL values for adducts from the cheek or tongue were not significantly different. Adduct levels in smokers (median RAL of 4.8 X 10(-8) were significantly higher (P less than 0.001) than adduct levels in non-smokers (median RAL of 2.9 X 10(-9). Adduct levels in drinkers (median RAL of 9.1 X 10(-10) were significantly lower (P less than 0.001) than adduct levels in non-drinkers (median RAL of 3.7 X 10(-8). Four of the subjects in this study have subsequently developed squamous cell carcinoma of the oral cavity. 32P-Postlabeling analysis of DNA from the oral cavity of these subjects did not demonstrate unique patterns or RAL values. Lack of information on the structure of the majority of adducts observed in this study was a serious limitation. Further improvements in adduct identification will be needed before 32P-postlabeling can be a useful tool for monitoring exposure of the oral cavity to carcinogens.

Topics: 7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; Adenosine Triphosphate; Alcohol Drinking; DNA; DNA Damage; DNA Replication; Humans; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Phosphorus Radioisotopes; Radioisotope Dilution Technique; Smoking; Tongue

Exfoliated mucosal cells were collected from the oral cavity of three groups at high risk for oral cancer: Indian betel nut chewers, Filipino inverted smokers (burning end of cigar in mouth) and Indian Khaini tobacco chewers. DNA was extracted from these samples, as well as from samples of exfoliated cells of Canadian non-smoking controls. DNA was analyzed for the presence of aromatic DNA adducts using 32P-postlabelling analysis. Five chromatographically distinct adducts were found in samples from both the high risk groups and the nonsmoking controls. Individual adducts were detectable in approximately 30-95% of samples, depending on the adduct and population group. Estimated levels of specific adducts ranged from non-detectable (prevalence relative to normal nucleotides less than 1 X 10(-9)) to occasionally greater than 1 X 10(-7). No adducts were found in high risk groups which did not also appear in control subjects.

Topics: Adenosine Triphosphate; Adult; Affinity Labels; Areca; Chromatography, High Pressure Liquid; Chromatography, Thin Layer; DNA; Female; Humans; Male; Middle Aged; Mouth Mucosa; Mouth Neoplasms; Phosphorus Radioisotopes; Plants, Medicinal; Plants, Toxic; Precancerous Conditions; Risk; Smoking; Tobacco, Smokeless

Biodistribution and tumor uptake studies were carried out with intravenously injected tracer doses of Gpp(NH)p labeled with 3H, 32P or 99mTc . Syrian golden hamsters with cheek pouch carcinomas, induced by repeated topical applications of DMBA, were used as a tumor model. The biodistributions of these three radionuclides were different, indicating significant molecular cleavage of this nucleotide analog. It was also apparent that this compound labeled with 99mTc may not be useful for tumor imaging due to low tumor-to-blood specific activity ratio. The cheek pouch carcinoma tumor model may be valuable for the evaluation of tumor localizing radiopharmaceuticals.

Topics: 9,10-Dimethyl-1,2-benzanthracene; Animals; Carcinoma, Squamous Cell; Cricetinae; Guanosine Triphosphate; Guanylyl Imidodiphosphate; Male; Mesocricetus; Mouth Neoplasms; Organotechnetium Compounds; Phosphorus Radioisotopes; Technetium; Tissue Distribution; Tritium

Topics: Adenoviridae; Autoradiography; Carbon Radioisotopes; Carcinoma; Cell Fractionation; Cell Line; Cell Nucleus; Choline; DNA Replication; DNA, Viral; Humans; Microscopy, Electron; Mouth Neoplasms; Phosphorus Radioisotopes; Thymidine; Thymine; Tritium

Topics: Adenosine Triphosphatases; Adenosine Triphosphate; Animals; Astrocytes; Carcinoma; Cell Line; Cell Membrane; Cell Membrane Permeability; Chlorine; Cricetinae; Diffusion; Dose-Response Relationship, Drug; Fibroblasts; HeLa Cells; Humans; Isotope Labeling; Mice; Mouth Neoplasms; Phosphorus Radioisotopes; Potassium Isotopes; Radioisotopes; Sodium Isotopes; Surface Properties

Topics: Base Sequence; Carcinoma; Cell Line; Cell Nucleus; Chromatography, Thin Layer; Diphosphates; DNA Nucleotidyltransferases; Electrophoresis, Paper; Electrophoresis, Polyacrylamide Gel; Escherichia coli; Exonucleases; Humans; Isoelectric Focusing; Kinetics; Magnesium; Manganese; Molecular Weight; Mouth Neoplasms; Phosphorus Radioisotopes; Templates, Genetic; Tritium

Purified particles of rhinovirus type 14 lose infectivity during incubation at 34.5 C as a result of fragmentation of RNA genomes within intact virions.

Topics: Carcinoma; Cell Line; Centrifugation, Density Gradient; Cesium; Chlorides; Electrophoresis, Polyacrylamide Gel; HeLa Cells; Humans; Molecular Weight; Mouth Neoplasms; Nucleic Acid Denaturation; Phosphorus Radioisotopes; Rhinovirus; RNA, Viral; Serotyping; Tritium; Uridine; Viral Plaque Assay

Topics: Animals; Autoradiography; Base Sequence; Carbon Radioisotopes; Carcinoma; Cell Line; Escherichia coli; Female; Formates; Humans; Hydrolysis; Methionine; Methylation; Mouth Neoplasms; Nucleosides; Perchlorates; Phosphorus Radioisotopes; Rats; RNA, Bacterial; RNA, Neoplasm; RNA, Transfer; Tritium; Uterus

Topics: Adenoviridae; Carcinoma; Cell Fractionation; Cell Line; Cell Nucleus; Centrifugation, Density Gradient; Cytoplasm; Dimethyl Sulfoxide; DNA, Viral; Electrophoresis, Polyacrylamide Gel; Helper Viruses; Humans; Molecular Weight; Mouth Neoplasms; Nucleic Acid Hybridization; Phosphorus Radioisotopes; Polyribosomes; RNA, Viral; Satellite Viruses; Sucrose; Transcription, Genetic; Tritium; Uridine

The RNA sequences and RNA size classes transcribed early in productive infection with adenovirus 2 were analyzed by RNA-DNA hybridization. Two independent procedures demonstrated that early cytoplasmic viral RNA is composed of two sequence classes, class I which is absent or present in greatly reduced quantities at 18 h, and class II which persists throughout the infection. When the sequences in early viral RNA were analyzed by hybridization-inhibition studies, the hybridization of early [(3)H]RNA was inhibited only 50% by RNA from cultures harvested late (18 h) in infection. Liquid hybridizations with radioactive viral DNA confirmed that early RNA includes two classes. Duplex formation of RNA with (32)P-labeled viral DNA was assayed by hydroxylapatite chromatography and resistance to S(1) nuclease digestion. Both methods showed that the cytoplasmic RNA present early in infection annealed 12 to 15% of the viral DNA; late cytoplasmic RNA hybridized 21 to 25% of the DNA. Mixtures of early plus late cytoplasmic RNAs hybridized 30 to 34% of the viral DNA, demonstrating the reduced concentration of early class I RNA in the late RNA preparations. Experiments were performed to correlate class I and class II early RNA with size-fractionated cytoplasmic RNA synthesized early in infection. Fractionation of RNA by gel electrophoresis or sucrose gradient centrifugation confirmed three major size classes, 12 to 15S, 19 to 20S, and 26S. Total cytoplasmic RNA and RNA selected on the basis of poly(A) content contained the same size classes of viral RNA. In standard electrophoresis conditions, the 19 to 20S viral RNA could be resolved into two size classes, and the distribution of 12 to 15S RNA also indicated the presence of more than one size component. Hybridization-inhibition studies under nonsaturating conditions were performed with 26S, 19 to 20S, and 12 to 15S viral RNAs fractionated by gel electrophoresis. Late RNA inhibited the hybridization of 26S RNA only 20%, 19 to 20S RNA was inhibited 45%, and 12 to 15S RNA was inhibited 50%. When 18 to 19S and 12 to 15S viral RNAs purified by sucrose gradient centrifugation were similarly analyzed, late RNA inhibited hybridization of 18 to 19S RNA 50%, and the annealing of 12 to 15S RNA was inhibited 70%.

Topics: Adenoviridae; Base Sequence; Carcinoma; Cell Line; Chromatography; Cytoplasm; DNA, Viral; Electrophoresis, Polyacrylamide Gel; Humans; Hydroxyapatites; Mouth Neoplasms; Nucleic Acid Hybridization; Phosphorus Radioisotopes; RNA, Viral; Time Factors; Transcription, Genetic; Tritium

Topics: Adenoviridae; Ammonium Sulfate; Carcinoma; Cell Line; Cell Nucleus; Cell-Free System; DNA-Directed RNA Polymerases; DNA, Viral; Humans; Isoenzymes; Mouth Neoplasms; Nucleic Acid Denaturation; Phosphorus Radioisotopes; Templates, Genetic; Transcription, Genetic

Topics: Acetates; Carcinoma; Cell Adhesion; Cell Line; Culture Techniques; Evaluation Studies as Topic; Glass; HeLa Cells; Leucine; Magnesium; Methods; Mouth Neoplasms; Phosphates; Phosphorus Radioisotopes; Tritium; Uridine

Topics: Adenoviridae; Caffeine; Carbon Radioisotopes; Carcinoma; Cell Line; Centrifugation, Density Gradient; Chromatography, DEAE-Cellulose; DNA Replication; DNA, Single-Stranded; DNA, Viral; Humans; Hydrogen-Ion Concentration; Isotope Labeling; Microscopy, Electron; Molecular Weight; Mouth Neoplasms; Nucleic Acid Hybridization; Phosphorus Radioisotopes; Thymidine; Time Factors; Tritium

Topics: Base Sequence; Carcinoma; Cell Line; Chromatography, Gel; DNA-Directed RNA Polymerases; Guanosine Triphosphate; Humans; Isoenzymes; Mouth Neoplasms; Phosphorus Radioisotopes; RNA; Transcription, Genetic

Topics: Adenoviridae; Caffeine; Carcinoma; Cell Line; Centrifugation, Density Gradient; Chromatography, Ion Exchange; Deoxyribonucleases; DNA Replication; DNA, Single-Stranded; DNA, Viral; Humans; Models, Chemical; Mouth Neoplasms; Neurospora crassa; Nucleic Acid Denaturation; Phosphorus Radioisotopes; Thymidine; Tritium; Virus Cultivation

Topics: Adenoviridae; Base Sequence; Carbon Radioisotopes; Carcinoma; Cell Line; Centrifugation, Density Gradient; DNA, Circular; DNA, Single-Stranded; DNA, Viral; Escherichia coli; Exonucleases; Humans; Microscopy, Electron; Molecular Weight; Mouth Neoplasms; Nucleic Acid Conformation; Nucleic Acid Denaturation; Nucleic Acid Renaturation; Phosphorus Radioisotopes; Satellite Viruses; Spectrophotometry, Ultraviolet; Thymidine; Tritium

Topics: Base Sequence; Carcinoma; Cell Line; Chlorella; Cytoplasm; Electrophoresis, Polyacrylamide Gel; Humans; Mouth Neoplasms; Phosphorus Radioisotopes; Ribonucleotides; RNA; Saccharomyces cerevisiae; Species Specificity

Topics: Adenoviridae; Autoradiography; Carcinoma; Cell Line; Centrifugation, Density Gradient; Cytarabine; Densitometry; Electrophoresis, Polyacrylamide Gel; HeLa Cells; Humans; Methionine; Microscopy, Electron; Molecular Weight; Mouth Neoplasms; Oxidative Phosphorylation; Peptide Biosynthesis; Peptides; Phosphates; Phosphorus Radioisotopes; Serotyping; Sulfur Radioisotopes; Viral Proteins; Virus Cultivation

Replicative intermediates of adenovirus type 5 DNA contain large stretches of single-stranded DNA. We have shown that this single-stranded DNA is mainly of parental origin, whereas all new DNA synthesized during one round of replication has a double-stranded structure. Hybridization experiments of the single-stranded DNA with isolated complementary strands of adenovirus type 5 DNA showed that this DNA hybridized only with the viral L-strand (the strand with the lower equilibrium density in alkaline CsCl) indicating that it represents the viral H-strand. This observation implies that replication always starts from one and the same molecular end. Electron microscopy of partially denatured Y-shaped intermediates confirmed this and showed that replication started from the molecular right end (the end richest in A-T base pairs). In conclusion, we have shown that replication of adenovirus type 5 DNA starts at the molecular right end, displacing the parental H-strand.

Topics: Adenosine Triphosphate; Adenoviridae; Carcinoma; Cell Line; Cell Nucleus; Centrifugation, Density Gradient; Cesium; Chromatography; DNA Replication; DNA, Single-Stranded; DNA, Viral; Humans; Microscopy, Electron; Mouth Neoplasms; Nucleic Acid Hybridization; Phosphorus Radioisotopes; Sucrose; Tritium; Virus Replication