phosphorus-radioisotopes and Liver-Neoplasms

PCBs are carcinogens, but for many decades it was assumed that PCBs may not possess initiating activity. Initiation is a process that involves changes in the DNA sequence, often, but not exclusively produced through DNA adduction by a reactive compound or reactive oxygen species (ROS). DNA adducts can be detected by (32)P-postlabeling, a method that Dr. Ramesh Gupta co-developed and refined. Today these types of assays together with other mechanistic studies provide convincing evidence that specific PCB congeners can be biotransformed to genotoxic and therefore potentially initiating metabolites. This review will provide an overview of our current knowledge of PCBs' genotoxic potential and mechanism of action, emphasizing the contributions of Dr. Ramesh Gupta during his tenures at the Universities of Kentucky and Louisville.

Topics: Animals; Carcinogens; Carcinoma, Hepatocellular; Cell Transformation, Neoplastic; DNA Adducts; DNA Damage; Humans; Isotope Labeling; Liver Neoplasms; Mutation; Phosphorus Radioisotopes; Polychlorinated Biphenyls; Toxicity Tests

Topics: Biocompatible Materials; Brachytherapy; Glass; Humans; Liver Neoplasms; Microspheres; Phosphorus Radioisotopes; Radiopharmaceuticals; Rhenium; Yttrium Radioisotopes

Magnetic resonance spectroscopy (MRS) remains the technique of choice for observing tumour metabolism non-invasively. Although initially 31P MR spectroscopy showed much promise as a non-invasive diagnostic tool, studies of a wide range of hepatic tumours have conclusively shown that this technique cannot be utilized to distinguish between different tumour types. This lack of specificity and sensitivity appears to be a consequence of the fact that hepatic tumours develop with a range of modalities and not as a single abnormal disease process, and also because of the limited availability of MR detectable metabolic markers. This has led, in recent years, to a re-evaluation of the role of 31P MR spectroscopy, re-emerging as a non-invasive tool to follow the efficacy of the treatment regime. Furthermore, since the principal changes observed in tumours by 31P MRS appear to be an elevation in the concentration of phosphorylcholine (PCho) and phosphoethanolamine (PEth), new research using a combination of MRS and tissue culture of cell lines which carry a combination of known inducible oncogenes, are helping to elucidate some of the metabolic pathways that give rise to these metabolic alterations.

Topics: Ethanolamines; Humans; Liver Neoplasms; Magnetic Resonance Spectroscopy; Phospholipids; Phosphorus Radioisotopes; Phosphorylcholine

Topics: Bone Neoplasms; Contrast Media; Germany; Humans; Iodine Radioisotopes; Leukemia, Radiation-Induced; Liver Neoplasms; Lung Neoplasms; Mining; Neoplasms, Radiation-Induced; Pacific Islands; Paranasal Sinus Neoplasms; Phosphorus Radioisotopes; Polycythemia Vera; Radioactive Fallout; Radiotherapy; Radium; Radon; Spondylitis, Ankylosing; Thorium; Thorium Dioxide; Thyroid Neoplasms; United States; Uranium

To evaluate the preventive effects of phosphorus-32 glass microspheres (P32-GMS) in the recurrence of massive hepatocellular carcinomas (HCCs) after tumor resection.. Twenty-nine patients with massive HCCs received local P32-GMS implantation after liver tumors were removed, while the other 38 patients with massive HCCs were not treated with P32-GMS after hepatectomies. The radioactivity of the blood, urine and liver were examined. The complications, HCC recurrence and overall survival rates in the patients were analyzed.. P32-GMS implanted in the liver did not cause systemic absorption of P32. There were no significant differences of postoperative complications between the patients with and without P32-GMS treatment. The short-term (six months and 1 year) and long-term (2, 3 and over 3 years) recurrence rates in patients who received P32-GMS radiotherapy were significantly decreased, and the overall survival rates in this group were significantly improved.. P32-GMS implantation in the liver can significantly decrease the postoperative recurrence and improve the overall survival in HCCs patients after hepatectomy. This therapy may provide an innovative method in prevention of HCC recurrence after operation.

Topics: Carcinoma, Hepatocellular; Combined Modality Therapy; Female; Glass; Hepatectomy; Humans; Liver Neoplasms; Male; Microspheres; Middle Aged; Neoplasm Recurrence, Local; Phosphorus Radioisotopes; Radiotherapy

The purpose of this study was to validate a previously reported body contour measurement using Compton backscatter sources with bremsstrahlung SPECT imaging.. Bremsstrahlung SPECT imaging was performed with 32P using a dual-headed camera system fitted with medium-energy, parallel-hole collimators. Two sources of 99mTc were placed directly on each collimator. Energy windows of 100 keV +/- 25% were used to image the 32P and also to record the Compton scatter from the 99mTc sources. Eleven patients enrolled in clinical Phase I therapeutic protocols were injected with 32P-chromic phosphate and SPECT images were acquired and reconstructed in the transaxial plane. The 32P distribution and the patient body contour were both visualized in these slices. The anteroposterior and lateral patient dimensions were measured by generating count profiles parallel to the anteroposterior and lateral body contour, respectively, at the midline in a transaxial slice. The distance in centimeters between the two centroids of each profile is representative of the anteroposterior and lateral dimensions and was determined for each patient. These anteroposterior and lateral dimensions were compared to the same distance measurements made in these patients by CT in an anatomically comparable transaxial slice. A cylindrical SPECT phantom was also studied to further validate the contour measurements.. The mean percent difference in the patient dimension measurements between SPECT and CT was -0.8% with a range of -8.5% to 9.9%. The percent difference between the known and SPECT measured dimensions in the cylindrical phantom was 0.5%.. The two external Compton scatter source method is accurate for determining the body contour.

Topics: Chromium Compounds; Head and Neck Neoplasms; Humans; Image Processing, Computer-Assisted; Liver Neoplasms; Lung Neoplasms; Pancreatic Neoplasms; Phantoms, Imaging; Phosphates; Phosphorus Radioisotopes; Scattering, Radiation; Technetium; Tomography, Emission-Computed, Single-Photon; Tomography, X-Ray Computed

In the past, we have clinically evaluated radiolabelled antibodies in Hodgkin's disease and hepatocellular cancer. Increased tumour pressure, reduced vascularity and poor diffusion has limited significant radiolabelled antibody tumour dose deposition. Using intratumoural infusion of macroaggregated albumin to blockade exiting vasculature followed by colloidal chromic 32Phosphorous, we have been able to achieve 75% to 100% tumour dose deposition by interstitial tumour infusion under computerised tomographic guidance. Phase I studies in a variety of solid tumours indicate extremely high doses may be achieved without toxicity (i.e. non-resectable pancreas 900,000 cGy to 1.7 million cGy) with tumour control and remission. This is a review of those studies and how the technique was applied.

Topics: Astrocytoma; Brachytherapy; Brain Neoplasms; Carcinoma, Hepatocellular; Carcinoma, Small Cell; Chemoembolization, Therapeutic; Chromium; Colloids; Dexamethasone; Head and Neck Neoplasms; Hodgkin Disease; Humans; Injections, Intralesional; Liver Neoplasms; Lung Neoplasms; Pancreatic Neoplasms; Phosphorus Radioisotopes; Radiography, Interventional; Radioimmunotherapy; Radiotherapy Dosage; Remission Induction; Serum Albumin; Tomography, X-Ray Computed

Radiation therapy plays a major role in the management of patients with either locally recurrent or metastatic carcinoma of the prostate.. In 23 patients with isolated postprostatectomy local recurrences treated with doses of 60-65 Gy, 17 (74%) had tumor control, and 45% survived relapse-free for 5 years after treatment of the recurrence. Pelvic irradiation has been used to treat patients with elevated prostate-specific antigen (PSA) levels after radical prostatectomy. This was tried, and 17 of 24 patients (70%) showed a significant decrease in PSA levels after irradiation, in five without subsequent elevation. Two of the seven patients with elevated PSA levels later had distant metastases. Local irradiation has been reported to yield excellent relief of symptoms in 100% of patients with hematuria, 80% with urinary outflow obstruction, and 50-70% with ureteral obstruction or pelvic pain secondary to locally advanced prostatic carcinoma. Reirradiation, particularly with brachytherapy (in preliminary studies combined with hyperthermia) has been used in the management of postirradiation prostatic recurrences with satisfactory tumor regression in approximately 75% of patients. The Radiation Therapy Oncology Group (RTOG) reported on the palliative effects of external irradiation on patients with bony metastasis. Approximately 54% of such patients had complete relief, and 29% had partial relief of bone pain. However, the retreatment rate of the bony metastasis was lower in the patients receiving higher doses. In a RTOG protocol in which all patients received local irradiation for osseous metastases, 77 were randomized to receive elective hemibody irradiation and 69, local treatment only. The frequency of additional treatment at 1 year was lower in the hemibody irradiation group (54% versus 78%). Occasionally, brain, mediastinal, or liver metastasis can be treated with irradiation. Radioactive phosphorus-32 or strontium-89 has been administered for disseminated bony metastasis with improvement of bone pain in approximately 70-80% of treated patients.. The role of irradiation in the treatment of spinal cord compression is discussed. Significant improvement of neurologic function has been reported in 36-60% of the patients, depending on severity of deficit and promptness in instituting emergency treatment.

Topics: Bone Neoplasms; Brachytherapy; Brain Neoplasms; Humans; Liver Neoplasms; Male; Neoplasm Recurrence, Local; Phosphorus Radioisotopes; Prostate-Specific Antigen; Prostatectomy; Prostatic Neoplasms; Radioisotopes; Radiotherapy Dosage; Rhenium; Spinal Cord Compression; Strontium Radioisotopes

The aim of this work is to evaluate the application of tissue-specific dose kernels instead of water dose kernels to improve the accuracy of patient-specific dosimetry by taking tissue heterogeneities into consideration.. Tissue-specific dose point kernels (DPKs) and dose voxel kernels (DVKs) for yttrium-90 (. The simulation results indicate that the highest differences between water and other tissue DPKs occur in bone for. A novel technique is proposed considering tissue-specific dose kernels in the dose calculation algorithm. This algorithm potentially enables patient-specific dosimetry and improves estimation of the average absorbed dose of

Topics: Algorithms; Bone Neoplasms; Computer Simulation; Humans; Liver Neoplasms; Lung Neoplasms; Lutetium; Monte Carlo Method; Organ Specificity; Phosphorus Radioisotopes; Radioisotopes; Radiometry; Radionuclide Imaging; Radiotherapy Planning, Computer-Assisted; Water; Yttrium Radioisotopes

The aim of this paper was to observe the metabolic mode of 32P at the level of sub-target nuclides.. Twenty-one cancer patients were locally injected with 32P-labelled glass microspheres and then observed to determine the equalization of 32P radionuclide metabolism in the tumor target. We imaged 3 sub-target regions of interest (ROI) 1/3 the size in both the anterior and posterior directions by bremsstrahlung single-photon emission computed tomography (SPECT) X-ray imaging. The radiation dose parameters of the beta rays including the initial dose rate, the effective half-life, and the effective half-life of the cumulative radiation dose were then calculated.. The radionuclide metabolism of the 21 complete tumor targets complied with the mono-compartmental model of index metabolism, but the level of tumor control did not correlate with radiation dose parameters. In contrast, the radionuclide metabolism of the 63 sub-targets did not comply with the mono-compartmental model. Instead, 32 sub-targets were better represented by bi-compartmental or tri-compartmental metabolic models. None of the remaining 31 sub-targets complied with index metabolism.. The complexity of the radiation dose at the sub-target level partially explains poor local tumor control. Future studies will be required to improve the expression of internal exposure to radiation dose parameters.

Topics: Adolescent; Adult; Aged; Carcinoma, Hepatocellular; Child; Female; Humans; Laryngeal Neoplasms; Liver Neoplasms; Lung Neoplasms; Lymphatic Metastasis; Male; Microspheres; Middle Aged; Neoplasms; Phosphorus Radioisotopes; Radiation Dosage; Rectal Neoplasms; Stomach Neoplasms; Tomography, Emission-Computed, Single-Photon; Young Adult

The objective of this study was to retrospectively evaluate the efficacy of a combination of (32)P-glass microsphere-mediated intra-arterial internal radiation and chemoembolization for the treatment of hepatocellular carcinoma.. Twenty-five consecutive patients with primary hepatocellular carcinoma referred for radiation therapy were treated with intra-arterial infusion of (32)P-glass microspheres followed by chemoembolization. beta-bremsstrahlung imaging was performed to monitor microsphere distribution. A partition model and a radiation dose equation were used for determination of radiation exposure in various tissues. Clinical response was evaluated using computed axial tomography scans.. The mean estimated absorption dose in tumor tissue was 137.42 +/- 56.69 Gy. A receiver operating characteristic curve was used to establish 90.65 Gy as the cutoff absorption dose with the best sensitivity and specificity for predicting response. The overall tumor response rate was 92%, while response in patients with radiation doses >90.65 Gy was 100%. Overall median patient survival was 15 months.. beta-bremsstrahlung imaging following intra-arterial infusion of (32)P-glass microspheres and chemoembolization incorporates effective treatment with convenient dosimetry monitoring and manageable adverse events using a single surgical procedure. This approach is a safe and effective method for ameliorating hepatocellular carcinoma.

Topics: Adult; Aged; Carcinoma, Hepatocellular; Chemoembolization, Therapeutic; Cohort Studies; Dose-Response Relationship, Radiation; Female; Humans; Infusions, Intra-Arterial; Kaplan-Meier Estimate; Liver Neoplasms; Male; Microspheres; Middle Aged; Neoplasm Invasiveness; Neoplasm Recurrence, Local; Neoplasm Staging; Phosphorus Radioisotopes; Radiography, Interventional; Radionuclide Imaging; Radiotherapy Dosage; Retrospective Studies; Risk Assessment; Survival Analysis; Tomography, X-Ray Computed

While potentially very useful, percutaneously delivered brachytherapy of inoperable intra-abdominal solid tumors faces significant technical challenges. This first-in-man study is designed to determine the safety profile and therapeutic efficacy of a novel phosphorous (32P) brachytherapy device (BrachySil) in patients with unresectable hepatocellular carcinoma.. Patients received single percutaneous and transperitoneal implantations of BrachySil under local anesthesia directly into liver tumors under ultrasound or computed tomographic guidance, at an activity level of 4 MBq/cc of tumor. Toxicity was assessed by the nature, incidence, and severity of adverse events (Common Toxicity Criteria scores) and by hematology and clinical chemistry parameters. Target tumor response was assessed with computed tomographic scans at 12 and 24 weeks postimplantation using World Health Organization criteria.. Implantations were successfully carried out in 8 patients (13-74 MBq, mean 40 MBq per tumor) awake and under local anesthesia. Six of the 8 patients reported 19 adverse events, but no serious events were attributable to the study device. Changes in hematology and clinical chemistry were similarly minimal and reflected progressive underlying hepatic disease. All targeted tumors were responding at 12 weeks, with complete response (100% regression) in three lesions. At the end of the study, there were two complete responses, two partial responses, three stable diseases, and one progressive disease.. Percutaneous implantation of this novel 32P brachytherapy device into hepatocellular carcinoma is safe and well tolerated. A significant degree of antitumor efficacy was demonstrated at this low dose that warrants further investigation.

Topics: Aged; Aged, 80 and over; Brachytherapy; Carcinoma, Hepatocellular; Female; Humans; Liver Neoplasms; Male; Middle Aged; Phosphorus Radioisotopes; Radiography, Interventional; Radiotherapy Dosage; Silicon Compounds; Tomography, X-Ray Computed; Treatment Outcome; Ultrasonography, Interventional

3-Nitrobenzanthrone (3-nitro-7H-benz[d,e]anthracen-7-one, 3-NBA) is a powerful mutagen and a suspected human carcinogen existing in diesel exhaust and airborne particulates. Recently, one of the major presumed metabolites of 3-NBA, 3-aminobenzanthrone (3-ABA), was detected in human urine samples. Here we analyzed DNA adducts formed in 3-NBA-exposed human hepatoma HepG2 cells by a (32)P-postlabeling/thin layer chromatography (TLC) method and a (32)P-postlabeling/polyacrylamide gel electrophoresis (PAGE) method. With HepG2 cells exposed to 3-NBA (0.36-36.4 microM) for 3h, we obtained three spots or bands corresponding to adducted nucleotides. Two were assigned as 2-(2'-deoxyadenosin-N(6)-yl)-3-aminobenzanthrone-3'-phosphate (dA3'p-N(6)-C2-ABA) and 2-(2'-deoxyguanosin-N(2)-yl)-3-aminobenzanthrone-3'-phosphate (dG3'p-N(2)-C2-ABA), with identical mobilities to those of synthetic standards on PAGE analysis. The chemical structure of the substance corresponding to the other spot or band could not be identified. Quantitative analyses revealed that the major adduct was dA3'p-N(6)-C2-ABA and its relative adduct labeling (RAL) value at 36.4 microM of 3-NBA was 200.8+/-86.1/10(8)nucleotide.

Topics: Benz(a)Anthracenes; Carcinoma, Hepatocellular; Cell Line, Tumor; Chromatography, Thin Layer; DNA Adducts; Electrophoresis, Polyacrylamide Gel; Humans; Liver Neoplasms; Phosphorus Radioisotopes

Topics: Adult; Carcinoma, Hepatocellular; Combined Modality Therapy; Embolization, Therapeutic; Female; Hepatic Artery; Humans; Injections, Intra-Arterial; Liver Neoplasms; Male; Microspheres; Middle Aged; Phosphorus Radioisotopes

Topics: Animals; Biological Transport; Carcinoma, Hepatocellular; Deoxyribonucleotides; Glycoconjugates; Humans; Ligands; Liver Neoplasms; Male; Metabolic Clearance Rate; Mice; Oligodeoxyribonucleotides, Antisense; Phosphorus Radioisotopes; Radioisotope Dilution Technique; Sulfur Radioisotopes; Thionucleotides; Tissue Distribution; Tumor Cells, Cultured

To evaluate the relationship between effects and internal radiation of phosphorus-32 glass microspheres embolization therapy for liver cancer patients.. From 1994 to 1998, 44 patients with unresectable liver cancer received intra-arterial radio-embolization therapy using (32)P-GMS. Preoperative and postoperative function and energy level of the liver were tested by liver function test and arterial blood ketone body ratio (AKBR). CT, single photon emission computer tomography (SPECT), and AFP were used to judge the effect of the therapy; multivariate analysis was made.. In the moderate dose group, low incidence of complication, high tumor shrinking rate and AFP decreasing rate, and long-term survival rate were observed. In the larger dose group, high incidence of liver failure, high tumor shrinking rate and AFP decreasing rate, and long-term survival rate were also observed. In the low dose group, low incidence of complication, but low tumor shrinking rate and AFP decreasing rate and long-term survival rate were not observed.. The reasonable radiation doses for liver cancer patients may be about 30 Gy; if liver cirrhosis is serious, the doses can be reduced.

Topics: Adult; Aged; Brachytherapy; Carcinoma, Hepatocellular; Embolization, Therapeutic; Female; Follow-Up Studies; Humans; Liver Neoplasms; Male; Microspheres; Middle Aged; Phosphorus Radioisotopes; Radiotherapy Dosage; Survival Rate

To investigate the efficacy of internal radiation of (32)P-glass microspheres ((32)P-GMS) in unresected hepatocellular carcinoma (HCC) via subcutaneous arterial port.. Hepatic arterial (99)technetium-macroaggregate albumin ((99)Tc-MAA) scanning via subcutaneous arterial port was undertaken to measure lung/liver shunting ratio and tumor/liver ratio. Hepatic arterial infusion of (32)P-GMS was performed in 17 cases of HCC with a dose from 1.11 to 1.30 GBq. Twenty cases of HCC undergoing hepatic arterial chemoembolization (HACE) in the same period served as controls group.. There was no treatment-related death in the 17 cases. In 7 of the 17 cases, AFP level and/or tumor size decreased by 50% after treatment, with a response rate of 64.7%. The median survival time was 5.5 months, and the 3-, 6-, 9-, 12-month survival rates were 94.1%, 44.1%, 31.0%, 24.4%, respectively. The therapeutic efficacy was better than that of HACE. The survival time was significantly longer in patients with T/N ratio >or= 2 than in those with T/N < 2 (P < 0.05).. Hepatic arterial infusion of (32)P-GMS is an alternative treatment for unresected HCC.

Topics: Adult; Aged; alpha-Fetoproteins; Carcinoma, Hepatocellular; Female; Follow-Up Studies; Hepatic Artery; Humans; Infusions, Intra-Arterial; Liver Neoplasms; Male; Microspheres; Middle Aged; Phosphorus Radioisotopes; Survival Rate

Infusional brachytherapy for treatment of neoplasms, with colloidal 32P has been used to treat various tumors in the pancreas, liver, brain, lung, and head and neck. In performing such treatments, anatomical verification of the location of the administered 32P from the image obtained by Bremsstrahlung SPECT alone is not possible due to the lack of internal landmarks, since the radionuclide is distributed only in the tumor and does not usually accumulate in the normal organs. The purpose of this study was to provide a practical three-dimensional approach for image fusion between Bremsstrahlung SPECT and CT.. The tumors in four cancer patients were injected directly with 32P under CT guidance. A Bremsstrahlung SPECT study using 99mTc backscatter sources to obtain the body contour was then performed. SPECT images were used to generate the skin contours using a threshold detection method. A three-dimensional surface was generated from these contours using a tiling program and fused with a corresponding CT surface generated from a CT scan in the same patient through an iterative surface-fitting algorithm. The three-dimensional surface of the region of high-activity, corresponding to the infused tumor, was then generated using the Bremsstrahlung SPECT data by mapping the iso-count surfaces through a computer program. The three-dimensional image of the organ then was fused with the registered CT-SPECT datasets.. The accuracy of fit measured as the mean distance between the SPECT and CT surfaces was in the range of 3-4 mm.. The anatomical co-registration of Bremsstrahlung SPECT with CT images using the outer surface-fitting algorithm is a reliable tool. This correlation permits direct anatomic confirmation of the region of the 32P activity distribution with the anatomic site selected for injection.

Topics: Algorithms; Brachytherapy; Humans; Kidney Neoplasms; Liver Neoplasms; Lung Neoplasms; Pancreatic Neoplasms; Phosphorus Radioisotopes; Radiotherapy Planning, Computer-Assisted; Tomography, Emission-Computed, Single-Photon; Tomography, X-Ray Computed

Parenchymal (PC) and nonparenchymal (NPC) liver cells have different tissue-specific, procarcinogen activation enzymes. NPC appear to be protected against the mutagenic effects of lipotropic bulky adducts induced by carcinogens by a unknown mechanism. Most studies of activation have been conducted with whole liver. The purpose of this study was to differentiate adduct formation in mouse PC and in NPC, isolated after in vivo administration of 7H-dibenzo(c,g)carbazole (DBC), the most efficient liver carcinogen in mice, which also has potent sarcomagenic and epitheliomagenic activities. The very sensitive 32P-postlabeling method was used to detect adducts. Two tissue-specific DBC derivatives, 6-methoxy-DBC (6MeODBC), which is exclusively sarcomagenic, and 5,9-dimethyl-DBC (DiMeDBC), which is exclusively hepatocarcinogenic, were analyzed in parallel. Both PC and NPC generated the ultimate metabolites of DBC, but NPC were substantially less efficient. Clear-cut tissue-specific differences in adduct formation were established: the sarcomagenic 6MeODBC gave rise only to NPC-DNA adducts, and the hepatocarcinogenic DiMeDBC only to PC-DNA adducts. The chromatograms of the adducts were compared with those of mouse embryonic cells in culture and mouse epidermal cells. The results are discussed in connection with animal experiments with DBC, 6MeODBC, and dimethylbenzanthracene and with published data on PC and NPC activating enzymes.

Topics: Animals; Carbazoles; Carcinogens; Centrifugation; DNA Adducts; Female; Isotope Labeling; Liver; Liver Neoplasms; Mice; Mice, Inbred Strains; Mice, Mutant Strains; Organ Specificity; Phosphorus Radioisotopes; Sarcoma, Experimental; Sensitivity and Specificity; Structure-Activity Relationship

Insulin binding to its receptor induces the phosphorylation of cytosolic substrates, insulin receptor substrate (IRS)-1 and IRS-2, which associate with several Src homology-2 domain-containing proteins. To identify unique IRS-1-binding proteins, we screened a human heart cDNA library with 32P-labeled recombinant IRS-1 and obtained two isoforms (epsilon and zeta) of the 14-3-3 protein family. 14-3-3 protein has been shown to associate with IRS-1 in L6 myotubes, HepG2 hepatoma cells, Chinese hamster ovary cells, and bovine brain tissue. IRS-2, a protein structurally similar to IRS-1, was also shown to form a complex with 14-3-3 protein using a baculovirus expression system. The amount of 14-3-3 protein associated with IRS-1 was not affected by insulin stimulation but was increased significantly by treatment with okadaic acid, a potent serine/threonine phosphatase inhibitor. Peptide inhibition experiments using phosphoserine-containing peptides of IRS-1 revealed that IRS-1 contains three putative binding sites for 14-3-3 protein (Ser-270, Ser-374, and Ser-641). Among these three, the motif around Ser-270 is located in the phosphotyrosine binding domain of IRS-1, which is responsible for the interaction with the insulin receptor. Indeed, a truncated mutant of IRS-1 consisting of only the phosphotyrosine binding domain retained the capacity to bind to 14-3-3 protein in vivo. Finally, the effect of 14-3-3 protein binding on the insulin-induced phosphorylation of IRS-1 was investigated. Phosphoamino acid analysis revealed that IRS-1 coimmunoprecipitated with anti-14-3-3 antibody to be weakly phosphorylated after insulin stimulation, on tyrosine as well as serine residues, compared with IRS-1 immunoprecipitated with anti-IRS-1 antibody. Thus, the association with 14-3-3 protein may play a role in the regulation of insulin sensitivity by interrupting the association between the insulin receptor and IRS-1.

Topics: 14-3-3 Proteins; Adenosine Triphosphate; Amino Acid Sequence; Animals; Binding Sites; Brain; Carcinoma, Hepatocellular; Cattle; Cell Line; CHO Cells; Cricetinae; Gene Library; Humans; Insulin; Insulin Receptor Substrate Proteins; Intracellular Signaling Peptides and Proteins; Liver Neoplasms; Mice; Molecular Sequence Data; Myocardium; Okadaic Acid; Phosphoproteins; Phosphorus Radioisotopes; Phosphotyrosine; Protein Biosynthesis; Proteins; Rats; Receptor, Insulin; Recombinant Proteins; Sequence Alignment; Sequence Homology, Amino Acid; Spodoptera; Transfection; Tumor Cells, Cultured; Tyrosine 3-Monooxygenase

The phosphorus-32 glass microspheres(32P-GMS) is a new radioembolizer for cancer therapy. From March 1994 to April 1995, 24 patients with unresectable hepatocellular carcinoma and 3 patients with cavernous hemangioma received internal radiation treatment of 32P-GMS. The clinical results demonstrated that hepatic arterial instillation of 32P-GMS appeared to be safe and effective for hepatocellular carcinoma. There was no significant bone marrow or renal toxicity or liver to pulmonary bypass. A transient change of liver function and fever occurred in almost all patients. Significant liver toxicity and gastrointestinal tract reaction were seen when radiation dose > 50 Gy or microspheres > 3 g, and then various postoperative complications occurred.

Topics: Adult; Brachytherapy; Carcinoma, Hepatocellular; Embolization, Therapeutic; Female; Fever; Glass; Hemangioma, Cavernous; Hepatitis; Humans; Liver Neoplasms; Male; Microspheres; Middle Aged; Phosphorus Radioisotopes; Radiation Dosage

Rats were treated with low doses of the hepatocarcinogens dimethylnitrosamine or thioacetamide, and livers were examined 48 h later. These treatments are known to produce altered RNA compartmentation, wherein a class of repetitive RNA sequences normally restricted to the nucleus appears in the cytoplasm. Reverse transcription-PCR amplifications demonstrated that the sequences showing altered compartmentation consisted largely of a subfamily of the rodent B2 sequence family, the counterpart of human Alu sequences involved in retrotransposition. Northern blot analyses showed that these B2 sequences were found in cytoplasmic RNA as 170- to 360-nucleotide "sense" transcripts, and competition hybridization experiments established that B2 sequences represented most (if not all) of the sequences showing altered compartmentation. The major increase in B2 transcriptions in cytoplasmic RNA was not associated with any change in B2 transcription by RNA polymerase III. In situ hybridizations showed that the altered compartmentation of B2 sequences occurred in well-delineated foci within the rat liver; these foci consisted of a central region containing a prominent infiltrate of macrophages admixed with small hepatocytes and a peripheral region of histologically normal hepatocytes that showed evidence of oxidative damage. Altered compartmentation of B2 sequences may represent an important focal initiatory change in a subset of hepatocytes, whereas subsequent retrotranspositional events (associated with Alu-like sequences) could predispose initiated cell foci to alterations in promotion/progression phases.

Topics: Animals; Base Sequence; Blotting, Northern; Carcinogens; Carcinoma, Hepatocellular; Cell Compartmentation; Dimethylnitrosamine; In Situ Hybridization; Liver Neoplasms; Male; Molecular Sequence Data; Nucleic Acid Conformation; Phosphorus Radioisotopes; Rats; Rats, Sprague-Dawley; Repetitive Sequences, Nucleic Acid; RNA, Antisense; RNA, Messenger; Thioacetamide; Time Factors

We evaluated the efficacy and side effect of 32P-Labelled glass microspheres (32P-GMS) as a radioembolizer for patients with advanced hepatocellular carcinoma (HCC). 24 patients with unresectable HCC received internal radiation treatment of 32P-GMS. The tumor size varied from 3.6 to 18 cm. Hepatic arterial embolization was carried out through intraoperative or Seldinger's method. The mean absorbed radiation dose of the liver was 3250 rad (range from 1200 rad to 8000 rad). The radiation intensity within the tumor was 3.3 times stronger than in liver tissue. Not significant bone marrow renal toxicity was noted within 1 to 3 months. > 50% of tumor shrinkage was found in 17 cases, and < 50% of tumor reduction in 5 cases. The cumulative survival rate of 3, 6, 12, 18, 24 months was 92%, 75%, 54%, 33% and 29%. Hepatic arterial instillation of 32P-GMS appears to be safe and effective for unresectable HCC even with portal vein thrombosis.

Topics: Adult; Brachytherapy; Carcinoma, Hepatocellular; Embolization, Therapeutic; Female; Glass; Humans; Liver Neoplasms; Male; Microspheres; Middle Aged; Phosphorus Radioisotopes; Survival Rate

Polychlorinated biphenyls (PCBs) are industrial chemicals which have been detected in fish, birds and humans. They are known to exert marked effects on the liver. They induce hepatocellular carcinoma in rats and birds, and are suspected of being carcinogenic to humans. To better understand the genotoxic effects of PCBs, we used 32P-postlabelling to investigate DNA adduct formation, after exposure to PCBs (Aroclor 1254 and 3,3',4,4'-tetrachlorobiphenyl), in primary cultures of fetal hepatocytes from two animal species and in a human cell line (Hep G2). We also studied the induction of 7-ethoxyresorufin-O-deethylase (EROD) in these PCB-treated cells. The three cell types used are known to express different cytochrome P450 families. The aim was to see whether a correlation could be established between EROD activity (a CYP1A1-related activity) and DNA adduct formation. DNA adducts were found in all three models after exposure to 50 microM 3,3',4,4'-tetrachlorobiphenyl. The number of adducts was higher in quail hepatocytes (37 adducts per 10(9) nucleotides) than in rat hepatocytes or Hep G2 cells (20 adducts per 10(9) nucleotides in both cases). The major adduct was the same in all three cell types, but some adducts were found in only one or two species. These inter-species differences probably reflect metabolic differences leading to different ultimate carcinogens. Exposure to Aroclor 1254 failed to produce significant levels of DNA adducts, suggesting that pre-treated cells are required to magnify Aroclor 1254 metabolism. No correlation was found between adduct formation and the level of EROD induction.

Topics: Analysis of Variance; Animals; Aroclors; Carcinogens, Environmental; Cells, Cultured; Chlorodiphenyl (54% Chlorine); Cytochrome P-450 CYP1A1; Cytochrome P-450 Enzyme System; DNA Adducts; Enzyme Induction; Hepatoblastoma; Humans; Liver; Liver Neoplasms; Oxidoreductases; Phosphorus Radioisotopes; Polychlorinated Biphenyls; Quail; Rats; Species Specificity; Tumor Cells, Cultured

Therapeutic agents such as monoclonal antibodies, radiopharmaceuticals, and radioactive growth factors are limited in effectiveness due to the inability to deposit significant quantities of the agents and for limited periods of time in solid cancer. A new technique based on knowledge of the pathophysiology of solid tumors allows for significant concentration of these agents to accumulate and for a prolonged period of time, thus allowing interaction with the tumor for potentially increased effectiveness.. Three agents have been studied: 131I antiferritin monoclonal antibody, colloidal 32P chromic phosphate, and 131I transferrin. The time required for maximal tumor uptake was determined in vitro in tissue culture and was 10 min, 25 min, and 40 min, respectively. The new method of in vivo tumor infusion consisted of a direct intratumoral injection of macroaggregated albumin (MAA) 10,000 particles, followed by the radioactive agents under study. Tumors were infused in vivo using the new technique and compared to intratumoral infused controls. In the instance of radiolabeled antiferritin antibody, intraperitoneal administration and intratumoral infusion were compared to the new technique. In the other two instances, intratumoral infusion was compared to the new method.. In all instances the direct vascular blockade caused by MAA led to greater deposition of the agent under study for at least 24 h. These results were clinically applied with MAA followed by 32P colloidal chromic phosphate and were consistent with the experimental findings.. A new technique is described that may be carried out in the experimental laboratory and clinic by direct tumor infusion of macroaggregated albumin (MAA), followed by other radioactive agents that will remain localized in solid cancers and will allow for high tumor dose deposition for potentially increased therapeutic efficacy.

Topics: Animals; Antibodies, Monoclonal; Antineoplastic Agents; Colloids; Cytotoxins; Ferritins; Humans; In Vitro Techniques; Iodine Radioisotopes; Liver Neoplasms; Mice; Mice, Nude; Phosphorus Radioisotopes; Rats; Transferrin; Tumor Cells, Cultured

The treatment of primary proliferative polycythaemia (polycythaemia rubra vera) may include radioactive phosphorus (P32) in conjunction with venesection. Acute leukaemia or carcinoma can be associated with the use of P32. We present a case of primary proliferative polycythaemia treated by repeat venesection together with P32 whose follow-up was complicated by the development of malignant neuroblastoma.

Topics: Bloodletting; Humans; Liver Neoplasms; Lung Neoplasms; Male; Middle Aged; Neuroblastoma; Phosphorus Radioisotopes; Polycythemia Vera

The asialoglycoprotein (ASGP) receptor undergoes constitutive endocytosis through the coated pit/coated vesicle pathway in hepatocytes. Studies on HepG2 cells have shown that the receptor is phosphorylated at serine under control conditions and following protein kinase C stimulation. This study examined whether the ASGP receptor could also serve as a substrate for a tyrosine kinase in HepG2 cells. 32P labeling was performed in membrane preparations, in permeabilized cells at 4 degrees C, and in intact cells at 37 degrees C. The phosphorylated ASGP receptor was isolated by immunoprecipitation, hydrolyzed in 6 N HCl at 110 degrees C, and analyzed by two-dimensional high voltage electrophoresis. The receptor isolated from a membrane preparation incubated in vitro with [gamma-32P]ATP incorporated radiolabel predominantly (greater than 90%) into phosphotyrosine. ASGP receptor phosphorylation at both tyrosine and serine was detected in intact cells incubated with phosphatase inhibitors for 60 min at 37 degrees C. The presence of both phenylarsine oxide (20 microM) and sodium orthovanadate (200 microM) was required for tyrosine phosphorylation. Use of these inhibitors together resulted in a 16.4-fold increase in phosphorylation of the immunoprecipitated ASGP receptor, whereas phosphorylation of total HepG2 membrane proteins was not significantly augmented by this procedure. Selective proteolytic digestion of ASGP receptors in isolated vesicles demonstrated that the phosphorylation site identified in these studies is located at tyrosine 5 in the cytoplasmic tail.

Topics: Asialoglycoprotein Receptor; Carcinoma, Hepatocellular; Cell Line; Cell Membrane; Endocytosis; Humans; Kinetics; Liver Neoplasms; Peptide Mapping; Phosphorus Radioisotopes; Phosphorylation; Receptors, Immunologic; Tyrosine

The levels of aromatic hydrocarbons in sediments of Puget Sound, Washington, are positively correlated with the prevalence of hepatic neoplasms and related lesions in English sole (Parophrys vetulus). To investigate the biochemical processes involved in chemical carcinogenesis in fish from Puget Sound, we have studied the uptake, activation, and detoxication of polycyclic aromatic hydrocarbons (PAHs) in English sole, and have compared these data to PAH metabolism in a related species, starry flounder (Platichthys stellatus), which shows a lower prevalence of hepatic neoplasms than sole. The results of both laboratory and field studies show that sediment-associated PAHs are biologically available to both flatfish species, and that both species accumulate similar levels of PAHs. Analyses of hepatic DNA from sole using the 32P-postlabeling technique indicate that xenobiotic chemicals were adducted to hepatic DNA of fish from the contaminated sites but not to the DNA of fish from reference sites. Studies of the ability of English sole and starry flounder to metabolize benzo(a)pyrene (BaP) and bind reactive BaP intermediates to hepatic DNA indicate that biochemical differences in the metabolism of carcinogenic PAHs may explain, at least in part, the apparent lower susceptibility of starry flounder than English sole to chemically induced hepatocarcinogenesis.

Topics: Animals; Benzo(a)pyrene; Biotransformation; DNA; Fish Diseases; Flatfishes; Flounder; Liver Neoplasms; Microsomes, Liver; Phosphorus Radioisotopes; Rats; Rats, Inbred Strains; Species Specificity; Washington; Water Pollutants; Water Pollutants, Chemical; Xenobiotics

The extent of embolization of different sized radioactive microspheres in experimental tumours and the homogeneity of their distribution in normal liver was examined in 25 rats. The ratio of arterially introduced microspheres lodging in tumour tissue compared to the surrounding normal hepatic parenchyma was measured for 15, 32.5 and 50 microns diameter tracer microspheres. The mean tumour to liver arterial perfusion ratio (T/L) for 15 and 32.5 microns spheres was approximately 3:1 in both cases and there was no significant difference between these values (P greater than 0.05). However, 50 microns microspheres did not preferentially lodge in malignant tissue (mean T/L ratio 1:1). The homogeneity of distribution of microspheres embolizing in the normal liver tissue was assessed for each microsphere size. As microsphere diameter increased from 15 to 50 microns, microspheres lodged more evenly throughout the liver substance. For 15 microns microspheres the coefficient of variation was 55.5% +/- 8.3 and 32.5 microns microspheres distributed with a coefficient of 35% +/- 16.8 while 50 microns spheres distributed most evenly with a coefficient of 19.7% +/- 6.8.

Topics: Animals; Kidney; Liver; Liver Neoplasms; Microspheres; Phosphorus Radioisotopes; Rats; Rats, Inbred Strains; Tissue Distribution; Yttrium Radioisotopes

Glass microspheres 20-30 microns in diameter containing either 31P or 89Y that can be activated by neutron bombardment to 32P or 90Y respectively, have been produced for intra-arterial radiotherapy of liver tumors. The spheres are insoluble in body fluids, non-toxic, and can concentrate in liver tumors of animals by direct injection into the hepatic artery. This gives a higher radiation dose to the tumor than can be achieved with external beam therapy. Trials in rabbits and dogs have proven successful and this product (90Y TheraSpheres) is now entering limited human clinical trials.

Topics: Animals; Dogs; Glass; Hepatic Artery; Humans; Injections, Intra-Arterial; Liver Neoplasms; Microspheres; Phosphorus Radioisotopes; Rabbits; Yttrium Radioisotopes

The bioenergetic state of 15 human tumours was examined with phosphorus-31 magnetic resonance spectroscopy. A striking diversity in metabolic patterns was observed, and significant differences from normal tissue were seen in all cases. A common feature was an elevation of intracellular pH, which may be related to an increase in Na+/H+ exchange during cell activation. It is unlikely that the patterns observed directly correlate with malignancy, but characterisation of the energetic state of a given tumour in a given physiological environment may help in the design and evaluation of interventions for that specific case.

Topics: Adult; Aged; Brain Neoplasms; Breast Neoplasms; Female; Humans; Hydrogen-Ion Concentration; Kinetics; Liver Neoplasms; Magnetic Resonance Spectroscopy; Male; Middle Aged; Neoplasms; Phosphorus Radioisotopes; Spectrum Analysis

Topics: Adrenal Gland Neoplasms; Biopsy; Ethanolamines; Female; Humans; Infant, Newborn; Liver; Liver Neoplasms; Magnetic Resonance Spectroscopy; Neuroblastoma; Phosphocreatine; Phosphorus Radioisotopes

Hepatocarcinogenic peroxisome proliferators, clofibrate, ciprofibrate, Wy-14643 or di(2-ethylhexyl)phthalate, were administered once daily by gavage to groups of three male F344 rats for 3 days and the rats were killed 2 h after the last dose. The DNA isolated from the livers was analyzed for possible carcinogen-DNA adducts, by a most sensitive 32P-postlabeling technique which can detect one adduct in 10(10) nucleotides. No adducts were detected by this assay in the DNA isolated from the livers of rats treated with any of the peroxisome proliferators. Adducts were also not found in the DNA of hepatocytes exposed in vitro to these peroxisome proliferators for 4 h in primary suspension cultures. Failure to detect peroxisome proliferator DNA adducts in hepatocytes under in vivo and in vitro conditions supports the contention that formation of a peroxisome proliferator-DNA adduct is not an essential step in the carcinogenesis by this novel class of carcinogens.

Topics: Animals; Anticholesteremic Agents; Carcinogens; Clofibrate; Clofibric Acid; Diethylhexyl Phthalate; DNA; Fibric Acids; Hypolipidemic Agents; In Vitro Techniques; Liver; Liver Neoplasms; Male; Microbodies; Phosphorus Radioisotopes; Pyrimidines; Rats

Commercial cation exchange resin beads (200--400 mesh) are separated into narrow particle size ranges by sieving and differential sedimentation. These microspheres are converted into the chromic form by reaction with Cr(NO2)3, labeled with 20 mCi 32P phosphate by exchange at pH 2--4, and converted to a stable product at pH 9. Chemical stability ahd biological behavior of these microspheres suspended in physiological saline is measured before administration through a hepatic artery catheter for radiation therapy of the liver and hepatic neoplasms.

Topics: Catheterization; Hepatic Artery; Humans; Ion Exchange Resins; Isotope Labeling; Liver Neoplasms; Microspheres; Phosphorus Radioisotopes; Quality Control

A 64-year-old patient underwent total abdominal hysterectomy and bilateral salpingo-oophorectomy in October, 1978 for a Stage IA, grade 2 papillary adenocarcinoma of the endometrium. Peritoneal washings contained numerous malignant cells, although the tumor invaded the myometrium only superficially. Two weeks after operation, 12 mCi of P32 were instilled into the peritoneal cavity. In May, 1979, laparotomy was performed for clinical obstruction of the small intestine and revealed diffuse peritoneal, omental, and hepatic metastases. Radiation changes involved the terminal ileum, ascending and sigmoid colon; an ileorectal fistula was also identified. The factors that might cause malignant cells to be present in the peritoneal cavity and the ideal treatment of such patients have yet to be determined. THe risk of intraperitoneal P32 might outweigh its benefits.

Topics: Adenocarcinoma, Papillary; Ascitic Fluid; Female; Humans; Liver Neoplasms; Middle Aged; Peritoneal Neoplasms; Phosphorus Radioisotopes; Uterine Neoplasms

Established cancer in the liver can, in selected patients who have a good arterial circulation in these tumors, be effectively treated by intrahepatic artery radioactive yttrium-90 resin microspheres. Even in unselected patients treated in the last five years by the author, 17 of 25 patients treated have had good objective regression of cancers, improvement of symptoms and prolongation of life. Treatment is relatively simple and associated with few side effects. For adjuvant therapy of colon cancer having positive nodes (Dukes C), internal radiation therapy of the liver is best done with Phosphorus-32 Colloid passed through the circulation of the gut to be effectively and homogeneously trapped by the Kupffer cells of the liver. Four such patients have been subjected to a pilot study--three of the four are doing well without significant side effects and no evidence of liver cancer after two years. When the fourth died of brain metastases, he too had less liver cancer than would be expected.

Topics: Aged; Animals; Brachytherapy; Colloids; Colonic Neoplasms; Female; Humans; Liver Neoplasms; Male; Microspheres; Middle Aged; Phosphorus Radioisotopes; Pilot Projects; Radiotherapy Dosage; Rats; Yttrium Radioisotopes

Purified nucleolar DNA was markedly degraded at a concentration of 13 mug/ml by bleomycin A2; bleomycin concentrations 20-30 times greater were required to degrade nucleoplasmic DNA. Whole nuclear DNA was degraded to only a small extent at 13 mug/ml but was markedly degraded at higher bleomycin concentrations. Treatment of the various types of DNA with high concentrations of bleomycin A2 produced low molecular weight (approximately 6S) fragments that were no longer sensitive to degradation by bleomycin A2. Hybridization studies demonstrated a loss of ribosomal DNA sequences from nucleolar DNA treated with bleomycin A2 in vitro. Studies on RNA synthesis in Novikoff hepatoma ascites cells in vitro showed there was a decreased uptake of 32Pi into high molecular weight nuclear RNA in the presence of bleomycin A2. These results indicate that nucleolar function is inhibited by a direct effect of bleomycin A2 on nucleolar DNA.

Topics: Ascitic Fluid; Bleomycin; Carbon Radioisotopes; Carcinoma, Hepatocellular; Cell Nucleolus; Cell Nucleus; Centrifugation, Density Gradient; DNA, Neoplasm; Leucine; Liver Neoplasms; Neoplasms, Experimental; Nucleic Acid Denaturation; Nucleic Acid Hybridization; Phosphorus; Phosphorus Radioisotopes; RNA, Neoplasm; RNA, Ribosomal; Thymidine; Transcription, Genetic; Tritium; Uridine

The phosphorylating of the lysine-rich histone at various stages in the cell cycle has been studied. In rapidly dividing cell populations the lysine-rich histone is phosphorylated rapidly after synthesis and more slowly once bound to the chromosome. The half-life of hydrolysis of such interphase phosphorylation in 5 hr except during mitosis when the phosphata hydrolysis increases almost three-fold. During mitosis there is extensive phosphorylation at sites different from those phosphorylated during interphase and a smaller measure of sites common to both mitotic and interphase cells. The sites of mitotic phosphorylation are most critically distinguished from those phosphorylated in interphase by the rapidly hydrolysis of M-phase phosphohistone when the cells divide and enter the G1 phase of the cell cycle.

Topics: Autoradiography; Carcinoma, Hepatocellular; Cells, Cultured; Demecolcine; DNA, Neoplasm; Electrophoresis; Histones; Hydroxyurea; Liver Neoplasms; Lysine; Mitosis; Neoplasms, Experimental; Phosphates; Phosphoproteins; Phosphoric Monoester Hydrolases; Phosphorus Radioisotopes; Tritium

Rat ascites hepatoma cell DNA polymerases (EC 2.7.7.7), especially low molecular weight polymerase, could incorporate a significant amount of single nucleotide into acid-soluble products in the absence of the other three deoxynucleoside triphosphates when activated DNA was used as a template. This relaxed requirement for deoxynucleotides was not observed when poly[d(A-T).d(T-A)] was used as a template. Nearest-neighbour base analyses of the products formed in the presence of a single deoxynuclesode triphosphate revealed that the reaction is not of a terminal transferase-type but a very limited repair synthesis in which one or a few triphosphates are incorporated at numerous 3'-hydroxyl ends.

Topics: Adenine Nucleotides; Adenosine Triphosphate; Animals; Carcinoma, Hepatocellular; Cells, Cultured; DNA; DNA Nucleotidyltransferases; DNA Repair; Liver Neoplasms; Methods; Molecular Weight; Nucleotides; Phosphorus Radioisotopes; Polynucleotides; Rats; Templates, Genetic

The capacity to effectively label tumor cell hostones using very short pulses of [3-H]acetate and [32-P]phosphate (1 to 10 min) has been developed. Four histone fractions F3, F2a1, F2a2, and F2b are extensively acetylated in short time periods. About 70% of the acetate accumulated on the histone during a short pulse is removed with a half-life of similar to 3 min. The rest of the metabolically active acetate is removed with a half-life of 30 to 40 min. Histones F2a1, F2a2, and F1 are acetylated at the NH2 terminus and this modification is metabolically stable. In short pulses, histones are labeled with 32-P in the order F2a2 greater than F1 greater than F3 greater than F2a1 greater than F2b. All fractions have a fairly rapid turnover time (t1/2 similar 20 to 40 min) except F1 phosphate which turns over some 5 times more slowly.

Topics: Acetates; Acetylation; Carcinoma, Hepatocellular; Cell Nucleus; Cells, Cultured; Cycloheximide; Histones; Liver Neoplasms; Neoplasms, Experimental; Phosphates; Phosphorus Radioisotopes; Tritium

After administration of N-nitrosomorpholine (NNM) to female Wistar-rats the ribosomal RNA (18 S and 28S rRNA) of the livers contained "abnormal" dinucleotides which were resistant against treatment with alkali or with spleen phosphodiesterase. These and further observations are discussed in view of the hypothesis that during the induction of liver tumors a metabolite of NNM causes crosslinks of nucleic acid bases. The application of this hypothesis on the effects of NNM upon DNA permits to explain the additional results that have been obtained. Observations on NNM metabolism as reported in the literature are not inconsistent with this interpretation.

Topics: Animals; Carcinogens; Female; Histocytochemistry; Liver Neoplasms; Morpholines; Nitroso Compounds; Nucleotides; Phosphoric Diester Hydrolases; Phosphorus Radioisotopes; Rats; RNA, Ribosomal

A variety of 6- and 8-substituted analogs of cAMP (cyclic adenosine 3:5-monophosphate) have been tested for their ability to increase activity of tyrosine aminotransferase (EC 2.6.1.5) in cultured Reuber H35 hepatoma cells. Some analogs, particularly the 8-thio-substituted ones, produced effects approximately equivalent to those generated by N-6, O2'-dibutyryl cAMP. In contrast, cAMP and its O-2-monobutyryl derivative were relatively ineffective even at very high concentrations, whereas three other analogs actually depressed the activity of the aminotransferase. Changes in enzyme activity generated by the various analogs were paralleled closely by changes in the relative rate of aminotransferase synthesis. An excellent correlation was found to exist between the ability of any given analog to influence the activity of tyrosine aminotransferase and that of phosphoenolpyruvate carboxykinase (EC 4.1.1.32). A similar correlation was found to exist between the ability of various analogs to evelate the activity of these enzymes and to inhibit reversibly the growth of H35 cells. Only one of five inhibitors of cAMP phosphodiesterase activity tested produce any increase in aminotransferase activity when added alone. All of the 6- and 8-substituted analogs tested, including noniducers, stimulated f1 histone phosphorylation in crude rat liver extracts with approximately equal potencies. On the other hand, dibutyryl cAMP was only a weak activator of protein kinase in vitro, even though it is a potent enzyme inducer. A possible resolution of this apparent discrepancy has been provided by preliminary analyses of site-specific f1 histone phosphorylation in whole cells. Only compounds active as aminotransferase inducers are capable of stimulating phosphorylation of the serine-37 residue of endogenous f1 histone (3- to 10-fold).

Topics: Animals; Butyrates; Carbon Radioisotopes; Carcinoma, Hepatocellular; Cell Line; Cyclic AMP; Cyclic GMP; Enzyme Activation; Inosine Nucleotides; Kinetics; Liver Neoplasms; Neoplasms, Experimental; Nucleotides, Cyclic; Phosphodiesterase Inhibitors; Phosphoenolpyruvate Carboxykinase (GTP); Phosphorus Radioisotopes; Protein Kinases; Rats; Structure-Activity Relationship; Theophylline; Time Factors; Tritium; Tyrosine Transaminase

Topics: Animals; Azo Compounds; Chromatin; Hepatectomy; Liver; Liver Neoplasms; Liver Regeneration; Male; Neoplasm Proteins; Neoplasms, Experimental; Nucleoproteins; Phosphates; Phosphoproteins; Phosphorus Radioisotopes; Protein Kinases; Rats; RNA; Templates, Genetic

Topics: Animals; Blood Cell Count; Bone Marrow Examination; Carcinoma 256, Walker; Colloids; Female; Injections, Intra-Arterial; Injections, Intravenous; Liver Neoplasms; Neoplasm Metastasis; Phosphorus Radioisotopes; Portal Vein; Rats

Topics: Alkaline Phosphatase; Animals; Carcinoma, Hepatocellular; Electrophoresis, Paper; Electrophoresis, Polyacrylamide Gel; Liver; Liver Neoplasms; Macromolecular Substances; Male; Neoplasms, Experimental; Organophosphorus Compounds; Phosphates; Phosphoproteins; Phosphorus Radioisotopes; Rats; Serine; Sodium Dodecyl Sulfate; Tyrosine Transaminase

Topics: Adenylyl Cyclases; Animals; Carcinoma, Hepatocellular; Dose-Response Relationship, Drug; Epinephrine; Fluorides; Glucagon; Iodine Radioisotopes; Liver; Liver Neoplasms; Male; Phosphorus Radioisotopes; Rats

Topics: Adenine Nucleotides; Animals; Base Sequence; Carcinoma, Hepatocellular; Chromatography, Gel; Electrophoresis, Starch Gel; Liver; Liver Neoplasms; Male; Neoplasms, Experimental; Phosphorus Radioisotopes; Polynucleotides; Polyribosomes; Rats; RNA, Neoplasm; RNA, Ribosomal; Sodium Dodecyl Sulfate

Topics: Animals; Base Sequence; Carcinoma, Hepatocellular; Cellulose; Chromatography, Ion Exchange; Chromatography, Paper; Cytosine Nucleotides; Electrophoresis; Electrophoresis, Paper; Hydrogen-Ion Concentration; Liver Neoplasms; Male; Neoplasms, Experimental; Phosphoric Diester Hydrolases; Phosphorus Radioisotopes; Polynucleotides; Pyrimidine Nucleotides; Rats; Ribonucleases; RNA, Ribosomal; Spleen; Uracil Nucleotides

Topics: Animals; Ascitic Fluid; Autoradiography; Base Sequence; Carcinoma, Hepatocellular; Chromatography, Ion Exchange; Coliphages; DNA Viruses; Electrophoresis, Polyacrylamide Gel; Hydrogen-Ion Concentration; Hydrolysis; Liver Neoplasms; Macromolecular Substances; Male; Neoplasms, Experimental; Nucleotides; Oligonucleotides; Pancreas; Phosphorus Radioisotopes; Polynucleotides; Pyrimidine Nucleotides; Rats; Ribonucleases; RNA, Neoplasm; RNA, Ribosomal

Topics: Adenosine Monophosphate; Animals; Carcinoma, Hepatocellular; Cell Nucleus; Chromatography, Ion Exchange; Cyclic GMP; Cytosine Nucleotides; Electrophoresis, Polyacrylamide Gel; Liver Neoplasms; Molecular Weight; Neoplasms, Experimental; Phosphorus Radioisotopes; Rats; Ribonucleotides; RNA; Uracil Nucleotides

Topics: Amino Acids; Animals; Ascitic Fluid; Autoradiography; Carcinoma, Hepatocellular; Cell Fractionation; Cell Nucleolus; Electrophoresis, Polyacrylamide Gel; Hydrogen-Ion Concentration; Liver Neoplasms; Male; Molecular Weight; Neoplasm Proteins; Neoplasms, Experimental; Organophosphorus Compounds; Peptide Fragments; Phosphates; Phosphoproteins; Phosphorus Radioisotopes; Pronase; Rats; Serine; Solubility; Threonine

Topics: Adenosine Triphosphatases; Animals; Ascitic Fluid; Carcinoma, Ehrlich Tumor; Carcinoma, Hepatocellular; Cell Fractionation; Dinitrophenols; Female; Hepatectomy; Leukemia L1210; Liver; Liver Neoplasms; Liver Regeneration; Male; Mice; Mitochondria, Liver; Neoplasms, Experimental; Oxidative Phosphorylation; Oxygen Consumption; Phosphates; Phosphorus Radioisotopes; Rats; Uncoupling Agents

Topics: Animals; Calcium; Carcinoma, Hepatocellular; Cell Fractionation; Cell Membrane; Lipoproteins; Liver; Liver Neoplasms; Magnesium; Neoplasms, Experimental; Phospholipids; Phosphorus Radioisotopes; Rats; RNA; RNA, Neoplasm

Topics: Base Sequence; Carcinoma, Hepatocellular; Cell Nucleolus; Chromatography, DEAE-Cellulose; Chromatography, Ion Exchange; Electrophoresis, Paper; Liver; Liver Neoplasms; Neoplasms, Experimental; Pancreas; Phosphorus Radioisotopes; Ribonucleases; Ribonucleotides; RNA; RNA, Ribosomal

Topics: Animals; Carbon Radioisotopes; Carcinoma, Hepatocellular; Cell Fractionation; Cell Nucleus; DNA, Neoplasm; Histones; Liver; Liver Neoplasms; Lysine; Male; Neoplasms, Experimental; Nucleoproteins; Phosphates; Phosphorus Radioisotopes; Radiation Effects; Rats; Rats, Inbred ACI; Thymidine; Time Factors

Topics: Animals; Carcinoma, Hepatocellular; Cell Nucleolus; Cell Nucleus; Centrifugation, Density Gradient; Electrophoresis, Polyacrylamide Gel; Liver Neoplasms; Neoplasms, Experimental; Nucleic Acid Hybridization; Phosphorus Radioisotopes; Rats; Ribonucleotides; RNA, Messenger; RNA, Neoplasm; RNA, Ribosomal; Spectrophotometry, Ultraviolet

Topics: Adenosine Triphosphate; Animals; Carcinoma, Hepatocellular; Cell Line; Cell-Free System; Chromatography, Thin Layer; Cyclic AMP; Cyclic GMP; Hot Temperature; Liver Neoplasms; Lymphoma; Pentosephosphates; Phosphorus Radioisotopes; Phosphotransferases; Protein Kinases; Rats; Ribose; Tritium

Topics: Adenosine Triphosphate; Animals; Carcinoma, Hepatocellular; Cell Fractionation; Cell Line; Chromatography, DEAE-Cellulose; Cyclic AMP; Cytoplasm; Cytosol; Dialysis; Drug Stability; Hydrogen-Ion Concentration; Liver; Liver Neoplasms; Male; Neoplasm Proteins; Neoplasms, Experimental; Phosphoric Diester Hydrolases; Phosphorus Radioisotopes; Protein Binding; Protein Kinases; Proteins; Rats; Tritium

Topics: Amoeba; Animals; Autoradiography; Carcinoma, Hepatocellular; Cell Line; Cell Nucleus; Chromatin; Cytoplasm; Dactinomycin; Densitometry; Electrophoresis, Polyacrylamide Gel; Liver Neoplasms; Molecular Weight; Phosphorus Radioisotopes; Rats; RNA; RNA, Neoplasm; Transcription, Genetic

Topics: Carcinoma, Hepatocellular; Cells, Cultured; Cycloheximide; DNA, Neoplasm; Enzyme Activation; Histones; Hydroxyurea; Kinetics; Liver Neoplasms; Lysine; Neoplasm Proteins; Neoplasms, Experimental; Phosphates; Phosphoric Monoester Hydrolases; Phosphorus Radioisotopes; Thymidine; Tritium

Topics: Animals; Base Sequence; Carcinoma, Hepatocellular; Centrifugation, Density Gradient; Chromatography, Ion Exchange; Chromatography, Paper; Electrophoresis, Paper; Electrophoresis, Polyacrylamide Gel; Hydrogen-Ion Concentration; Hydrolysis; Liver Neoplasms; Male; Molecular Weight; Neoplasms, Experimental; Pancreas; Phosphoric Diester Hydrolases; Phosphorus Radioisotopes; Rats; Ribonucleases; Ribonucleotides; Ribosomes; RNA, Neoplasm; RNA, Ribosomal; Snakes; Spleen; Time Factors; Venoms

Topics: Adenosine Triphosphate; Animals; Calcium; Carcinoma, Hepatocellular; Cell Nucleolus; Cobalt; Cyclic AMP; Electrophoresis, Disc; Electrophoresis, Polyacrylamide Gel; Hydrogen-Ion Concentration; Kinetics; Liver Neoplasms; Magnesium; Manganese; Neoplasm Proteins; Neoplasms, Experimental; Phosphoproteins; Phosphorus Radioisotopes; Protein Kinases; Rats; Solubility; Time Factors; Zinc

Topics: Animals; Base Sequence; Carcinoma, Hepatocellular; Cell Nucleus; Cellulose; Centrifugation, Density Gradient; Chromatography, DEAE-Cellulose; Chromatography, Ion Exchange; Chromatography, Paper; Electrophoresis; Electrophoresis, Paper; Electrophoresis, Polyacrylamide Gel; Liver Neoplasms; Neoplasms, Experimental; Oligonucleotides; Pancreas; Phosphorus Radioisotopes; Rats; Ribonucleases; Ribonucleotides; RNA, Neoplasm

Topics: Acetates; Animals; Ascites; Carcinoma, Hepatocellular; Cell Nucleolus; Centrifugation, Density Gradient; Deoxyribonucleases; Edetic Acid; Electrophoresis; Electrophoresis, Polyacrylamide Gel; Histocytochemistry; Kinetics; Liver Neoplasms; Magnesium; Male; Microscopy, Electron; Neoplasms, Experimental; Nucleotides; Pepsin A; Phosphates; Phosphorus Radioisotopes; Polyvinyls; Proteins; Rats; Ribonucleases; Ribosomes; RNA, Ribosomal; Sucrose

Topics: Animals; Base Sequence; Carcinoma, Hepatocellular; Cellulose; Chromatography, Ion Exchange; Chromatography, Paper; Drug Stability; Electrophoresis; Electrophoresis, Paper; Electrophoresis, Polyacrylamide Gel; Hydrogen-Ion Concentration; Liver Neoplasms; Molecular Weight; Neoplasms, Experimental; Oligonucleotides; Pancreas; Phosphorus Radioisotopes; Rats; Ribonucleases; Ribonucleotides; RNA, Neoplasm

Topics: Animals; Carcinoma, Hepatocellular; Cell Nucleolus; Cell Nucleus; Chromatin; Electrophoresis, Polyacrylamide Gel; Isotope Labeling; Liver Neoplasms; Neoplasm Proteins; Neoplasms, Experimental; Phosphoproteins; Phosphorus Radioisotopes; Rats; Ribosomes; Serine; Time Factors

Topics: Animals; Carcinoma, Hepatocellular; Cyclic AMP; Enzyme Activation; Isoelectric Focusing; Isoenzymes; Liver; Liver Neoplasms; Neoplasms, Experimental; Phosphorus Radioisotopes; Protein Kinases; Rats; Ultracentrifugation

Topics: Adenosine Triphosphate; Amino Acids; Animals; Ascitic Fluid; Carbon Radioisotopes; Carcinoma, Hepatocellular; Cell Nucleolus; Centrifugation, Density Gradient; Cytoplasm; Diphosphates; Hydroxamic Acids; Hydroxylamines; Liver Neoplasms; Neoplasm Proteins; Neoplasms, Experimental; Phosphates; Phosphorus Radioisotopes; Rats; RNA, Transfer; Transfer RNA Aminoacylation

Topics: Animals; Calcium; Carcinoma, Hepatocellular; Lanthanum; Leukemia, Experimental; Liver Neoplasms; Lymphoma, Non-Hodgkin; Magnesium; Melanoma; Mice; Neoplasms, Experimental; Phosphates; Phosphorus Radioisotopes; Sarcoma, Experimental; Sodium

Topics: Animals; Carcinoma, Hepatocellular; Cations, Divalent; Cations, Monovalent; Cell Nucleolus; Centrifugation, Density Gradient; Chromatography, Ion Exchange; Chromatography, Paper; Coliphages; Electrophoresis, Polyacrylamide Gel; Endonucleases; Hydrogen-Ion Concentration; Kinetics; Liver Neoplasms; Macromolecular Substances; Male; Molecular Weight; Neoplasms, Experimental; Nucleoproteins; Pancreas; Phosphorus Radioisotopes; Rats; Ribonucleases; RNA, Ribosomal; Spectrophotometry, Ultraviolet; Temperature

Topics: Animals; Carbon Radioisotopes; Carcinoma, Hepatocellular; Chromatography, Gel; Cytosol; Leucyl Aminopeptidase; Liver; Liver Neoplasms; Neoplasm Proteins; Neoplasms, Experimental; Phosphorus; Phosphorus Radioisotopes; Protein Binding; Proteins; Rats

Topics: Animals; Carcinoma, Hepatocellular; Liver; Liver Neoplasms; Male; Neoplasms, Experimental; Nucleic Acid Hybridization; Nucleotides; Oligonucleotides; Phosphorus Radioisotopes; Rats; RNA, Neoplasm; RNA, Ribosomal

Topics: Carcinoma, Hepatocellular; Cell Nucleus; Centrifugation, Density Gradient; Deoxyribonucleotides; Electrophoresis; Hot Temperature; Hydrogen-Ion Concentration; Liver Neoplasms; Neoplasms, Experimental; Nucleic Acid Conformation; Osmolar Concentration; Phosphorus Radioisotopes; RNA; RNA, Ribosomal; Sodium Chloride; Spectrophotometry, Ultraviolet; Temperature

Topics: Animals; Ascites; Carcinoma, Hepatocellular; Cell Membrane; Glycerophosphates; Liver; Liver Neoplasms; Lysophosphatidylcholines; Male; Molecular Weight; Neoplasms, Experimental; Nucleic Acids; Phosphatidylethanolamines; Phosphatidylinositols; Phospholipids; Phosphorus Radioisotopes; Rats; Sphingomyelins

DNA was extracted from [(3)H]thymidine-labeled Marek's disease virus (MDV) and purified by two cycles of CsCl gradient centrifugation in a fixed-angle rotor. The DNA was transcribed in vitro into (32)P-labeled complementary RNA (cRNA). MDV cRNA did not hybridize with DNA from chicken embryo fibroblast cultures or from chicken spleen, but hybridized efficiently with DNA from MDV particles or MDV-infected cell cultures. Five Marek's disease tumors from different chickens and different organs (ovary, liver, testis) were all found to contain MDV DNA sequences. The relative amount of MDV DNA varied from tumor to tumor and was between 3 and 15 virus genome equivalents per cell. The content of virus DNA per cell in spleens from tumor-bearing chickens was much lower than in tumors from the same animals. MDV-infected cell cultures contained a large proportion (28-59%) of virus antigen-positive cells, as measured by immunofluorescence, but tumor cells were negative in this respect (<0.02% positive cells). These data indicate that MDV is present in a provirus form in tumor cells.

Topics: Animals; Base Sequence; Centrifugation, Density Gradient; Chick Embryo; DNA, Viral; Ducks; Embryo, Nonmammalian; Female; Fibroblasts; Fluorescent Antibody Technique; Herpesviridae; Liver Neoplasms; Male; Marek Disease; Microscopy, Electron; Neoplasms, Experimental; Nucleic Acid Hybridization; Ovarian Neoplasms; Phosphorus Radioisotopes; RNA, Viral; Testicular Neoplasms; Tritium; Virus Cultivation

Topics: Animals; Ascitic Fluid; Base Sequence; Carcinoma, Hepatocellular; Cell Nucleolus; Cell Nucleus; Cells, Cultured; Cellulose; Centrifugation, Density Gradient; Electrophoresis; Electrophoresis, Paper; Electrophoresis, Polyacrylamide Gel; Hydrogen-Ion Concentration; Liver Neoplasms; Male; Molecular Weight; Neoplasms, Experimental; Nucleic Acid Conformation; Oligonucleotides; Phosphorus Radioisotopes; Rats; Ribonucleotides; RNA; RNA, Ribosomal

Topics: Coloring Agents; Liver Neoplasms; Mechlorethamine; Nitrogen Mustard Compounds; Phosphorus; Phosphorus Radioisotopes; Pyridines; Pyridinium Compounds

Topics: Animals; Azo Compounds; Carcinogens; Coloring Agents; Cytoplasm; Liver Neoplasms; Neoplasms; Phosphorus; Phosphorus Radioisotopes; Rats