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F F Becker

Publications and source records attributed to F F Becker.

At least 91 records · Page 5Linked to original sources

Tumor phenotype and susceptibility to progression as an expression of subpopulations of initiated murine cells.

Currently, it is conceived that a number of events, or hits, are required for the induction of tumors by chemical agents. The first phase of this sequence, initiation, is considered to result from at least one event in the genetic apparatus. Analyses of this sequence, however, usually give little consideration to the nature of the target cell or to the characteristics of the resultant tumors. Vesselinovitch et al. (Cancer Res., 38: 2003-2010, 1978) have reported that a single, small pulse of carcinogen can induce early and numerous liver tumors when administered neonatally to mice with a genetic predisposition to hepatotumorigenesis. In the current study, the nonpredisposed strain C57BL/6N was also shown to be highly susceptible to diethylnitrosamine during the neonatal period. C57BL/6N demonstrated large numbers of two of the three types of liver tumors seen in livers of genetically predisposed mice, one of which required the additional stimulus of dietary phenobarbital for growth. Tumors of more malignant phenotype were demonstrated only in genetically predisposed mice (C57BL/6N X C3H/HeN F1) that received one dose of carcinogen. These findings suggest that the phenotype of a tumor that results from a pulse of a chemical carcinogen may depend upon the target cell. The initiated cells that result from this hit may vary from those that demonstrate very little progression in cell type and may or may not require exogenous enhancement of growth to those that can progress very rapidly to fully malignant behavior. The latter might arise from a hit in a genetically initiated cell, the result of which is a more rapid progression in tumor type.

Animals↗

Proviral structure and differentiation antigen phenotype of spontaneous and chemically induced AKR lymphomas.

AKR mice develop spontaneous thymomas after 6 months of age due to a novel class of murine leukemia viruses that are generated by a series of genetic recombinations between endogenous proviral loci. AKR mice also are more susceptible to N-methyl-N-nitrosourea (MNU)-induced thymomas than are low-leukemia-incidence mouse strains. To determine whether virally and chemically induced lymphomagenesis proceeds by similar pathways in AKR mice, spontaneous and MNU-induced thymic lymphomas were analyzed for a DNA restriction linkage generated during spontaneous tumor development by recombination between envelope genes of endogenous murine leukemia proviral loci. DNA from spontaneous thymic lymphomas invariably contained a restriction fragment characteristic of recombinant murine leukemia virus etiology, while four of five MNU-induced thymic lymphomas did not show this restriction linkage. In addition, analysis of lymphocyte differentiation antigen profiles indicated that MNU-induced lymphomas represent a more immature stage of T-cell differentiation than the majority of spontaneous lymphomas. These data suggest that there are fundamental differences in the mechanisms of induction of virally and chemically induced thymic lymphomas in AKR mice.

Animals↗

Mechanism of rat liver DNA methyltransferase interaction with anti-benzo[a]pyrenediol epoxide modified DNA templates.

We investigated the methylation reaction catalyzed by 1500-fold purified rat liver DNA methyltransferase (DMase) on native Micrococcal luteus DNA (ML-DNA) and poly(dC-dG) templates containing covalently bound (+)-7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (anti-BPDE), the strongly carcinogenic, principal metabolite of benzo[a]pyrene. Since eukaryotic DNA methyltransferases recognize the dinucleotide 5'd[CG] in DNA as a substrate for methylation, the model polynucleotide poly(dC-dG) was used to study in more detail the mode of interaction and effect on incorporation. With either of these BPDE-modified templates, a progressive inhibition of methylation was correlated with increasing amount of BPDE substitution. The effect of BPDE-dG adducts did not alter the apparent km with respect to the concentration of d[CG] in either unmodified or BPDE-modified poly(dC-dG) (km = 10 microM) but lowered the relative apparent Vmax. In assays in which perturbation by salt of preformed enzyme-DNA complex is measured, no change in the relative stability to either unsubstituted or the carcinogen-modified template was noted, thus, excluding any change in the ionic component of this interaction. However, in competition-type experiments, BPDE-DNA is an inhibitor of the methylation reaction on native DNA. When BPDE-DNA is allowed to interact with the enzyme before the addition of native competitor DNA, the methylation rate is not stimulated, suggesting very tight hydrophobic binding of the enzyme to BPDE-DNA and an inhibition in the dissociation of DMase from the template following a methylation event.(ABSTRACT TRUNCATED AT 250 WORDS)

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗

Interaction of DNA methyltransferase with aminofluorene and N-acetylaminofluorene modified poly(dC-dG).

Poly(dC-dG) was reacted in vitro to yield templates containing similar amounts of aminofluorene (AF) or acetylaminofluorene (AAF) adducts. These modified poly(dC-dG) templates were tested in an in vitro DNA methylation system utilizing 1500-fold purified rat liver methyltransferase (DMase) to compare and quantitate the effects of these adducts on the kinetics of methylation and the interaction of DMase with such templates. Enzymatic methylation is severely impaired by arylamine adducts, with bound AF inhibiting more than AAF (relative Vmax 0.24 for AAF-poly(dC-dG) and 0.066 for AF-poly(dC-dG). The apparent km for the reaction is not appreciably altered by AAF modification: 10 microM for dCdG dinucleotide units, but it is threefold lower (3 microM) for AF-poly(dC-dG). In competition experiments it was demonstrated that a translocational block is imposed by the adducts. From differential salt inhibition assays and preincubation assays, no change in the ionic binding to the altered templates could be detected, which suggests that the enzyme interacts very strongly through hydrophobic interactions with the fluorene ring. Evidence that the fluorene ring is exposed is supported by circular dichroism spectra of the templates under the conditions of the assay, which indicated that the AF adducts do not appreciably change the normal B conformation of the template, while the template with 9.5% modification by AAF adducts adopted a Z form. These results suggest that the inhibitory effects of AAF and, in particular, AF upon DMase-catalyzed methylation reactions are not dependent upon helix conformation. Instead, they appear to depend upon DMase recognition of an altered dG base configuration, which is responsible for altered binding and methylation kinetics.

2-Acetylaminofluorene↗

Temperature-dependent transmembrane potential changes in cells infected with a temperature-sensitive Moloney sarcoma virus.

Normal rat kidney cells (NRK) infected with the temperature-sensitive (ts) transformation mutant of Moloney murine sarcoma virus yielded a clone of cells, 6m2, that exhibited a transformed morphology at 33 degrees C and a normal morphology at 39 degrees C. Transmembrane potential (Em) was measured fluorometrically using a cyanine dye diS-C3-(5). Fluorescence was inversely correlated with Em. Cells at 33 degrees C had lower Em. Em changes were recorded within 15 minutes of temperature shift from 33 degrees C to 39 degrees C in both directions, increasing in the 33 degrees C to 39 degrees C direction and decreasing in the 39 degrees C to 33 degrees C direction. Uninfected NRK cells when shifted under the same condition exhibited small fluorescence changes in the 33 degrees C to 39 degrees C direction. Shifting from 39 degrees C to 33 degrees C resulted in Em changes similar to those in 6m2 cells. Also studied was a cell line infected with a spontaneous revertant of the ts mutant, designated 54-5A4; it was transformed at both temperatures. Shifting from 33 degrees C to 39 degrees C in both directions yielded small changes. Transmembrane potential changes in 6m2 cells precede other transformation-specific changes that occur after a temperature shift.

Animals↗

A constitutive deficiency in the monooxygenase system of spontaneous mouse liver tumors.

Exposure to chemical carcinogens evokes a population of altered hepatocytes that demonstrates significantly diminished monooxygenase activity. It has been suggested that this alteration permits the target cell to escape the toxic effects of the carcinogen and proliferate. In an attempt to determine whether this enzyme defect has broader implications for the carcinogenic process, we examined the monooxygenase system and additional components of spontaneous hepatocellular tumors in mice with a genetic predisposition to tumorigenesis. These tumors uniformly demonstrated a significant deficit in cytochrome P-450 and aminopyrine N-demethylase, despite the absence of known carcinogens, toxins, or promoting agents in their environment. Tumors of similar histiotype induced by a small, single neonatal administration of diethylnitrosamine demonstrated identical alterations. This report, therefore, suggests a strong link between a genetic program for tumorigenesis and a deficit in the monooxygenase system in spontaneous tumors. Further, it reveals that a toxic-selective environment is not required for the expansion of the cell population that possesses this phenotype.

Animals↗

Methylation of the alpha-fetoprotein gene in productive and nonproductive rat hepatocellular carcinomas.

The extent of methylation of Hpall-Mspl and Hhal sites in DNAs isolated from normal rat livers and from the transplantable hepatocellular carcinomas (THC) THC 7777 and THC 252 was compared. It was found that the overall level of methylation of the internal cytosine in CCGG sequences was lower in the THC DNAs than in the normal liver DNAs. This difference could also be detected in the extent of methylation of CCGG sites flanking a 400-base pair repetitive sequence. Examination of methylation of specific sites within the alpha-fetoprotein gene revealed differences between the DNAs that appear to reflect both the level of activity of the gene and the overall level of methylation of cellular DNA. This gene, which is repressed in normal adult liver and the nonproductive THC 252 and highly active in the productive THC 7777 (S. Sell et al., Cell Biol. Int. Rep., 4: 235-254, 1980), contains several CCGG sites that are methylated in both normal liver and THC 252 DNA but not in THC 7777 DNA. However, Hhal (GCGC) sites in the alpha-fetoprotein gene were less methylated in both hepatoma DNAs than in liver DNA, which the exception of one site in the productive tumor found to be no longer methylated.

Animals↗

Circumoral chemical peel combined with cervicofacial rhytidectomy.

Forty consecutive cervicofacial rhytidectomies were performed; 36 were carried out in women and four in men, giving a rate of 10% for male rhytidectomies. Thirty-one of the female patients and one male patient had a circumoral chemical peel performed the day of surgery or the following day for a rate of 80% circumoral peel combined with rhytidectomy. The cases were carefully reviewed for results, and it was believed that circumoral chemical peeling combined with rhytidectomy greatly enhanced the overall result in the majority of cases in which it was performed. The only complication noted was slight hypopigmentation of the peeled skin compared with the surrounding cheek skin in some cases, which is easily covered with cosmetics and did not present a problem in any of the cases.

Aging↗

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Chemexfoliation↗

Immunocytochemical localization of gamma-glutamyltransferase in induced hyperplastic nodules of rat liver.

The immunocytochemical localization of gamma-glutamyltransferase [(5-glutamyl)-peptide:amino-acid 5-glutamyltransferase, EC 2.3.2.2; gamma-GluTase] was demonstrated in hyperplastic liver induced by the carcinogen 2-acetylaminofluorene (2-AAF). The method used a specific antiserum and protein A-horseradish peroxidase and permitted visualization of antigenic sites at both the light and electron microscopic levels. Electron microscopy revealed deposits of 3,3'-diaminobenzidine (DAB) reaction product in the plasma membranes of (i) hyperplastic cells, (ii) bile canaliculi, (iii) endothelial cell membranes, and (iv) lymphocytes. The so-called ATPase activity was localized in the plasma membrane in bile canaliculi and in endothelial cells; the hyperplastic cells show marked variability in the levels of this activity.

2-Acetylaminofluorene↗

The protective role of thiol reducing agents in the in vitro inhibition of rat liver DNA methylase by direct acting carcinogens.

The direct-acting carcinogens N-acetoxy-N-acetyl-2-aminofluorene (AcAAF), methyl nitrosourea (MNU), and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) were tested for their ability to inhibit highly purified, rat liver DNA methylase in vitro. Fifty percent inhibition of DNA methylase activity was achieved with 4.3 mM AcAAF, 47 mM MNU and 2.8 mM MNNG. When the enzyme was reassayed in the presence and absence of dithiothreitol, it was shown that DNA methylase was protected by increasing amounts of the thiol reducing agent. When other thiol reducing agents were tested for their ability to protect DNA methylase from carcinogen damage, a differential protective ability was observed. Dithiothreitol, beta-mercaptoethanol, and reduced glutathione were effective in protecting DNA methylase from carcinogen inhibition, while the effect of cysteine was intermediary and the effect of ergothioneine was minimal. These results may be related to the hypomethylation of DNA observed in several cancers, suggesting that the carcinogens achieve this effect at least in part by inhibiting crucial sulfhydryl group(s) in the methylase molecule. These data also suggest that various intracellular thiols may play an important role in protecting DNA-modifying enzymes from carcinogen damage.

Acetoxyacetylaminofluorene↗

The direct and indirect effects of promoters may depend upon the nature of the initiated cell.

The sequence of events in carcinogenesis may be more complex than previously proposed. We cannot as yet define the nature of initiation, nor in fact the obligate aspects of promotion. Administering small doses of diethylnitrosamine (DEN) to neonatal mice genetically predisposed to hepatocarcinogenesis, or to lines that are not (with and without subsequent phenobarbital (PB) administration), gave evidence that different types of initiated cells might exist. The majority of tumors that resulted were identical in morphology and biology to those that arise spontaneously from basophilic, diploid cells. However, a second tumor type was seen only in DEN-PB treated mice. This was composed of huge eosinophilic, polyploid cells. A third tumor type, rapidly progressing to a fully malignant form, was seen only in predisposed mice that received DEN, suggesting a genetic "hit" in a genetically predisposed cell. In addition to demonstrating the spectrum of initiated cells available for the action of promoters, evidence was obtained that indicated that one major action of PB was to stimulate growth of these altered cell populations rather than to induce new phenotypic alterations commensurate with progression. Thus, characteristics of PB-expanded, altered hepatocyte populations could be identified in very early cell foci.

2-Acetylaminofluorene↗

Thioacetamide hepatocarcinogenesis.

Thioacetamide (TAA) was administered to inbred male ACI rats for 1 year to determine its hepatocarcinogenicity. The carcinogen was fed at a level of 0.035% in a semipurified diet so that other components could be manipulated. The following groups were used: group I, TAA alone; group II, TAA with riboflavin content at 0.002% (one-third of the daily requirement); group III, TAA plus a supplement of 0.05% crude bovine thyroid; and group IV, TAA, 0.002% riboflavin, and thyroid supplement. All groups demonstrated cirrhosis, neoplastic nodules, and cholangiofibromas to varying degrees. Group IV was invariably the most advanced in all lesions, while group II demonstrated the next most advanced picture of cholangiofibromas. These lesions persisted for 1 year after cessation of the diet. Primary hepatocarcinomas (PHC) were produced by groups I, III, and IV, with the highest number being in group IV. Group II evoked only primary cholangiocarcinomas (PCC). A number of both types of carcinomas transplanted successfully. Thus TAA appears to be a hepatocarcinogen capable of inducing either PHC or PCC, with the tumor type evoked somewhat dependent on the associated diet.

Acetamides↗

Occurrence of progressive DNA damage coincident with the appearance of foci of altered hepatocytes.

The technique of alkaline elution was used to evaluate alterations in nuclear DNA obtained from livers of rats that had received a single 6.6 mg/kg dose of diethylnitrosamine (DEN) during liver regeneration and subsequent exposure (7 days after partial hepatectomy) to phenobarbital (BP). DNA from normal and regenerating liver demonstrated a significant increase in the rate of elution following DEN administration. In those DEN-treated groups that did not receive PB, the rate of DNA elution decreased slowly but failed to return to normal by 44 weeks. Exposure to PB hastened recovery of a normal DNA elution profile in normal liver following DEN treatment, by 44 weeks, DNA from these rats eluted at a normal rate. However, in rats treated with DEN during liver regeneration, the rate of DNA elution began to increase at 28 weeks of PB exposure and became progressively more rapid through the 36th and 44th weeks. This latter group also demonstrated foci of gamma-glutamyl-transferase (GGT)-positive hepatocytes at 28 weeks of PB exposure that increased in number and size concomitantly with the increasing rate of DNA elution. AT 44 weeks, one or more primary hepatocellular carcinomas were present in 73% of the rats in this group; none was seen in any other group. Foci of GGT positive hepatocytes, an increasing rate of DNA elution and eventual primary hepatocellular carcinoma were seen in a group of rats that was begun on PB as late as 85 days after partial hepatectomy and DEN treatment.

Animals↗

Hepatocellular ligandin during N-2-fluorenylacetamide carcinogenesis.

Ligandin was decreased by 75% as determined immunologically and by glutathione-S-transferase or steroid isomerase activities in rat hepatocellular carcinomas induced by exposure to N-2-fluorenylacetamide. Minor variable differences in ligandin levels were noted between the putative, premalignant nodules induced by this regimen and normal liver.

2-Acetylaminofluorene↗