Does a diet rich in dietary fibre really reduce the risk of colon cancer?
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Colonic Neoplasms↗
Biomedical subjects
L R Ferguson
Publications and source records attributed to L R Ferguson.
Mechanisms by which resistant starches and non-starch polysaccharide sources affect the metabolism and disposition of the food carcinogen, 2-amino-3-methylimidazo[4,5-f]quinoline.
Although both non-starch polysaccharides (NSP) and resistant starches (RS) are included in current definitions of dietary fibre, our previous work has suggested fundamental differences in the way in which these two classes of material affect the disposition and absorption of a dietary carcinogen. The present studies explore whether different effects on carcinogen metabolism could play a role in the contrasting patterns seen previously. Groups of female Wistar rats were pre-fed for 4 weeks one of five types of defined diet (AIN-76). The control diet contained 35% maize starch and no dietary fibre. The RS-containing diets had all the maize starch substituted with either Hi-maize or potato starch. In the NSP-containing diets, 10% of the maize starch was substituted with dietary fibre in the form of either lignified plant cell walls (wheat straw) or soluble dietary fibre (apple pectin). Pre-fed rats were gavaged with the food carcinogen, [2-14C] 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), and plasma and urinary metabolites characterized using HPLC at various time intervals after administration. After 4 h gavage, plasma from rats on both RS-containing diets contained significantly higher levels of intact IQ and lower levels of the major metabolites, IQ-5-O-glucuronide and IQ-5-sulfate, as compared with plasma from the negative control group at this time. In contrast, plasma from animals on the NSP-containing wheat straw diet (and to a lesser extent the apple pectin diet) showed significantly lower levels of intact IQ, and significantly higher levels of the two major metabolites, as compared with those from the control rats. These different metabolite profiles were also reflected in different urinary excretion profiles. Urine from rats pre-fed RS-containing diets revealed significantly slower metabolite excretion as compared with urine from rats that had been given the NSP-containing diets. Western blotting methodologies also profiled differences between the effects of these two types of dietary fibre in the expression of xenobiotic metabolizing enzymes. We conclude that changes in activity and expression of xenobiotic metabolising enzymes could play a role in the contrasting effects of these two types of dietary fibre on carcinogen uptake and disposition.
Animals↗
The 10 basic requirements for a scientific paper reporting antioxidant, antimutagenic or anticarcinogenic potential of test substances in in vitro experiments and animal studies in vivo.
There is increasing evidence that chemicals/test substances cannot only have adverse effects, but that there are many substances that can (also) have a beneficial effect on health. As this journal regularly publishes papers in this area and has every intention in continuing to do so in the near future, it has become essential that studies reported in this journal reflect an adequate level of scientific scrutiny. Therefore a set of essential characteristics of studies has been defined. These basic requirements are default properties rather than non-negotiables: deviations are possible and useful, provided they can be justified on scientific grounds. The 10 basic requirements for a scientific paper reporting antioxidant, antimutagenic or anticarcinogenic potential of test substances in in vitro experiments and animal studies in vivo concern the following areas: (1) Hypothesis-driven study design; (2) The nature of the test substance; (3) Valid and invalid test systems; (4) The selection of dose levels and gender; (5) Reversal of the effects induced by oxidants, carcinogens and mutagens; (6) Route of administration; (7) Number and validity of test variables; (8) Repeatability and reproducibility; (9) Statistics; and (10) Quality Assurance.
Animals↗
Antimutagenesis/anticarcinogenesis 2001: screening, methods and biomarkers.
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Animals↗
Antimutagenesis/anticarcinogenesis 2001: mechanistic studies.
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Animals↗
Vitamins/minerals and genomic stability in humans.
Recommended dietary allowances (RDAs) of micronutrients have been traditionally derived as those levels necessary to prevent symptoms of deficiency diseases. There is increasing evidence that higher levels of many such micronutrients may be necessary for various DNA maintenance reactions, and that the current RDAs for some micronutrients may be inadequate to protect against genomic instability. Supplementation of a normal diet, with either vitamins and/or minerals or with isolated plant polyphenols, is becoming increasingly common in most Western populations. However, there is no clear agreement as to how much supplementation should occur, if at all, and genotypic differences are not accounted for. The 14 mini-reviews in this special issue summarise the role of specific micronutrients in various aspects of DNA maintenance: DNA synthesis, DNA repair, DNA methylation, gene mutation, chromosome breakage, chromosome segregation, gene expression, oxidative stress, necrosis and apoptosis. Evidence has been collated from mammalian and human experiments, both using in vitro cultures and in vivo approaches. Authors were asked to critically assess the strength of evidence as to whether the micronutrient can affect genomic stability in humans at realistic intake levels, and to estimate optimal dietary ranges where possible. Information on further research necessary is also documented. These reviews are an essential step towards a definition of RDAs designed to maintain genomic stability.
DNA↗
Role of plant polyphenols in genomic stability.
Polyphenols are a large and diverse class of compounds, many of which occur naturally in a range of food plants. The flavonoids are the largest and best-studied group of these. A range of plant polyphenols are either being actively developed or currently sold as dietary supplements and/or herbal remedies. Although, these compounds play no known role in nutrition (non-nutrients), many of them have properties including antioxidant, anti-mutagenic, anti-oestrogenic, anti-carcinogenic and anti-inflammatory effects that might potentially be beneficial in preventing disease and protecting the stability of the genome. However not all polyphenols and not all actions of individual polyphenols are necessarily beneficial. Some have mutagenic and/or pro-oxidant effects, as well as interfering with essential biochemical pathways including topoisomerase enzyme activities, prostanoid biosynthesis and signal transduction. There is a very large amount of in vitro data available, but far fewer animal studies, and these are not necessarily predictive of human effects because of differences in bacterial and hepatic metabolism of polyphenols between species. Epidemiological studies suggest that high green tea consumption in the Japanese population and moderate red wine consumption in the French population may be beneficial for heart disease and cancer, and these effects may relate to specific polyphenols. A small number of adequately controlled human intervention studies suggest that some, but not all polyphenol extracts or high polyphenol diets may lead to transitory changes in the antioxidative capacity of plasma in humans. However, none of these studies have adequately considered long-term effects on DNA or the chromosome and unequivocally associated these with polyphenol uptake. Furthermore, clinical trials have required intravenously administered polyphenols at concentrations around 1400mg/m(2) before effects are seen. These plasma concentrations are unlikely to be achieved using the dietary supplements currently available. More focused human studies are necessary before recommending specific polyphenolic supplements at specific doses in the human population.
Animals↗
Flow cytometric enumeration of micronucleated reticulocytes: high transferability among 14 laboratories.
This laboratory previously described a single-laser flow cytometric method, which effectively resolves micronucleated erythrocyte populations in rodent peripheral blood samples. Even so, the rarity and variable size of micronuclei make it difficult to configure instrument settings consistently and define analysis regions rationally to enumerate the cell populations of interest. Murine erythrocytes from animals infected with the malaria parasite Plasmodium berghei contain a high prevalence of erythrocytes with a uniform DNA content. This biological model for micronucleated erythrocytes offers a means by which the micronucleus analysis regions can be rationally defined, and a means for controlling interexperimental variation. The experiments described herein were performed to extend these studies by testing whether malaria-infected erythrocytes could also be used to enhance the transferability of the method, as well as control intra- and interlaboratory variation. For these studies, blood samples from mice infected with malaria, or treated with vehicle or the clastogen methyl methanesulfonate, were fixed and shipped to collaborating laboratories for analysis. After configuring instrumentation parameters and guiding the position of analysis regions with the malaria-infected blood samples, micronucleated reticulocyte frequencies were measured (20,000 reticulocytes per sample). To evaluate both intra- and interlaboratory variation, five replicates were analyzed per day, and these analyses were repeated on up to five separate days. The data of 14 laboratories presented herein indicate that transferability of this flow cytometric technique is high when instrumentation is guided by the biological standard Plasmodium berghei.
Animals↗
Contrasting effects of acute and chronic dietary exposure to 2-amino-3-methyl-imidazo[4,5-f]quinoline (IQ) on xenobiotic metabolising enzymes in the male Fischer 344 rat: implications for chemoprevention studies.
BACKGROUND: 2-Amino-3-methyl-imidazo[4,5-f]quinoline (IQ) is a mutagen produced in cooked food. It is commonly present in the human diet and often used as a (pro)carcinogen for chemoprevention studies. Many foodstuffs act as chemopreventers by altering xenobiotic metabolising enzyme expression in favour of detoxication over bioactivation pathways. However, IQ itself can also affect enzyme expression, which may be a confounding factor in chemoprevention studies. AIM OF STUDY: Chronic low dose IQ exposure is intuitively closest to the human dietary situation. The aim was to investigate the effects of chronic dietary exposure to IQ on the expression of enzymes involved in the bioactivation and detoxification of xenobiotics and to compare this with acute exposure, often used in chemoprevention studies. METHODS: Male Fischer rats received IQ (300 ppm) in the diet (AIN-76) for 52 weeks or were given IQ (20 mg.kg-1) orally for 3 days. Animals were killed, livers removed and subcellular fractions prepared. A range of enzymes was selected to allow investigation of several cellular mechanisms. Enzyme expression and activity were determined by Western blotting and the use of selective probe substrates as appropriate. RESULTS: Chronic exposure to IQ led to an increase in phase II detoxifying enzymes. Both the activity and expression of glutathione S-transferase (GST-A1/2) were increased, as were NADPH: Quinone oxidoreductase (NQO), UDP-glucuronosyl transferase (UGT) and beta-glucuronidase activities. There were no statistically significant changes in the potential for bioactivation by three cytochrome P450s. In contrast, acute IQ exposure significantly increased the expression and activity of some cytochrome P450 (CYP1A1 and CYP1A2), UGT and beta-glucuronidase, but significantly decreased glutathione S-transferase expression and activity. There was a non-significant decrease in NQO but no change in CYP3A2 and CYP2E1 activities. CONCLUSIONS: The changes after acute exposure suggest an interaction through the Ah receptor and xenobiotic response element, modified by the glucocorticoid response element. In contrast, the pattern of effects after chronic exposure suggests activation of the antioxidant response element (ARE). Although the acute model is more practically convenient for short-term chemoprevention screening, the data suggest that an entirely new mechanism is being invoked that completely masks effects of the ARE that occur during chronic exposure. There is a danger that chemopreventive strategies developed using acute models may be misleading, since the mechanism is unlikely to occur during human dietary exposures.
Animals↗
Evidence for epidermal growth factor receptor-enhanced chemosensitivity in combinations of cisplatin and the new irreversible tyrosine kinase inhibitor CI-1033.
Irreversible inhibitors of the epidermal growth factor receptor (EGFR) are showing promise in clinical trials. This report is the first to show that inhibition of the EGFR tyrosine kinase by an irreversible binder synergizes with cisplatin, at least in EGFR-overexpressing tissue culture cell lines in vitro. Unlike previous synergies demonstrated between ErbB2 blockade and DNA-damaging drugs, the synergy between the irreversible EGFR inhibitor and cisplatin does not appear to involve the repair of DNA-cisplatin adducts. Given the current clinical data, this combination may be of more than theoretical interest.
Animals↗
Changing concepts of dietary fiber: implications for carcinogenesis.
The dietary fiber (DF) hypothesis suggested that DF, as plant cell walls, protected against colorectal cancer. The implicit assumption in much historic literature was that the readily analyzed and quantified nonstarch polysaccharide (NSP) component was critical in cancer protection. However, the presence of polymeric phenolic components such as lignin or suberin has profound effects on the physicochemical properties of the cell walls and largely determines their physiological properties in humans. In certain groups of food plants, degradation of cell walls that contain neither lignin nor suberin releases ferulic acid and other hydroxycinnamic acids. These acids have antioxidant, antimutagenic, and other anticancer effects, including modulation of gene expression and immune response. Reexamination of literature on cancer protection suggests that plant cell walls containing significant amounts of phenolic components may be the most likely to protect against cancer. In the last 30 years, the definition of DF has been incrementally expanded to incorporate NSP extracted from plant cell walls and from sources other than plant cell walls, then resistant starch, and most recently nondigestible oligosaccharides. On the early definitions, increased consumption of DF could only be achieved by increasing the intake of whole grains, brans, or other food plant material that would increase the intake of plant cell walls containing significant amounts of phenolic components. However, the new definitions make it possible to increase "dietary fiber" without consuming any such materials. We suggest that this could have negative connotations for cancer risk in human populations.
Animals↗
HUman MicroNucleus project: international database comparison for results with the cytokinesis-block micronucleus assay in human lymphocytes: I. Effect of laboratory protocol, scoring criteria, and host factors on the frequency of micronuclei.
Micronucleus (MN) expression in peripheral blood lymphocytes is well established as a standard method for monitoring chromosome damage in human populations. The first results of an analysis of pooled data from laboratories using the cytokinesis-block micronucleus (CBMN) assay and participating in the HUMN (HUman MicroNucleus project) international collaborative study are presented. The effects of laboratory protocol, scoring criteria, and host factors on baseline micronucleated binucleate cell (MNC) frequency are evaluated, and a reference range of "normal" values against which future studies may be compared is provided. Primary data from historical records were submitted by 25 laboratories distributed in 16 countries. This resulted in a database of nearly 7000 subjects. Potentially significant differences were present in the methods used by participating laboratories, such as in the type of culture medium, the concentration of cytochalasin-B, the percentage of fetal calf serum, and in the culture method. Differences in criteria for scoring micronuclei were also evident. The overall median MNC frequency in nonexposed (i.e., normal) subjects was 6.5 per thousand and the interquartile range was between 3 and 12 per thousand. An increase in MNC frequency with age was evident in all but two laboratories. The effect of gender, although not so evident in all databases, was also present, with females having a 19% higher level of MNC frequency (95% confidence interval: 14-24%). Statistical analyses were performed using random-effects models for correlated data. Our best model, which included exposure to genotoxic factors, host factors, methods, and scoring criteria, explained 75% of the total variance, with the largest contribution attributable to laboratory methods.
Adolescent↗
Antimutagenic effects of wheat bran diet through modification of xenobiotic metabolising enzymes.
Diets containing wheat bran (WB) protect against cancers of the colon or breast in rats, and may be beneficial in humans. In a previous study of rats treated with the carcinogen 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), inclusion of 10% wheat bran in the diet led to an apparent reduction in IQ metabolites but not of intact IQ in plasma. In the present study, male Wistar rats were fed diets containing 0, 10 or 20% wheat bran, and effects on xenobiotic metabolising enzymes compared. Wheat bran-supplementation showed differential effects on phase I enzymes, significantly increasing the activity of hepatic cytochrome P450 isozyme CYP3A2, but slightly reducing the activity of CYP1A1/2. The activities of both hepatic phase II detoxification enzymes glutathione-S-transferase and glucuronosyl transferase were also reduced. Western blotting revealed similar effects on expression of the proteins. Interestingly, the expression of xenobiotic metabolising enzymes (XME) in the colon appeared to be modulated independently of hepatic XME. Although the wheat bran-supplemented diet still led to an increased expression of CYP3A, it now slightly increased CYP1A in the colon. However, 20% wheat bran significantly increased the expression of both glutathione transferase isozymes, GST A1 & A2, in the colon. Natures Gold (NG) is a commercial wheat bran derivative which is lower than wheat bran in dietary fibre, but enriched in vitamins, minerals and various phytochemicals. Dietary supplementation with 20% Natures Gold led to similar trends as seen in wheat bran-fed rats, but more potent effects in both hepatic and colonic enzymes. The significance of these changes for activation of carcinogens to mutagenic metabolites was investigated using the Salmonella/mammalian microsome mutagenicity test. The activation of IQ and benzo[a]pyrene, but not cyclophosphamide, to a mutagen by hepatic S9 from wheat bran-fed or Natures Gold-fed rats was significantly reduced compared with S9 from animals on a diet lacking wheat bran. We suggest that modulation of xenobiotic metabolising enzymes may be an important component of cancer protection by wheat bran, and this effect may relate to micronutrients or cancer-protective non-nutrient phytochemicals rather more than to dietary fibre.
Animals↗
The mutagenic spectrum of acridine-linked aniline nitrogen mustards in AS52 cells: implications of DNA targeting with high selectivity for adenine or guanine bases.
The mutational spectra generated in AS52 cells at the gpt gene locus by aniline mustards were studied by the isolation of resistant clones and sequencing of the altered gene. A set of four aniline mustards (both mono- and bifunctional) linked to a DNA-affinic 9-aminoacridine (9-AA) carrier was used, together with the untargeted mustards chlorambucil (CHL) and its half-mustard, and the DNA binding carrier, 9-AA. Both 9-AA and CHL were weak cytotoxins, with the DNA-targeted mustards being markedly (10-40-fold) more dose potent, and the bifunctional ones somewhat more toxic than the monofunctional ones. 9-AA produced a different spectrum of mutations to the spontaneous background, with more minor addition events and less base pair substitutions, and showing for the first time that frameshift events so characteristic of 9-AA in bacteria or bacteriophage also occur in mammalian cells. The mutational spectra of the DNA-targeted mustards were quite different both from this and from the lesions caused by the untargeted mustards, which cause largely transition mutations at AT sites (despite a clear preference for formation of N(7)-guanine adducts). There were very few transition mutations, suggesting that the initial O(6)-alkylguanine/O(4)-alkylthymine lesions considered to give rise to these are relatively rare. There was also a lower incidence of complete deletions, usually attributed to DNA cross-links. For the short chain length targeted mustards, which form initial stable adducts largely (95%) at guanine N(7) sites, base pair substitution mutations, predominantly transversions, involved AT and GC base pairs equally. In contrast, the longer chain length targeted mustards, which form >90% of initial adducts at adenine N(1) sites, generated also formed transversion mutations, but these overwhelmingly (24/27) involved AT base pairs.
Aminacrine↗
Transcriptional blockages in a cell-free system by sequence-selective DNA alkylating agents.
There is considerable interest in DNA sequence-selective DNA-binding drugs as potential inhibitors of gene expression. Five compounds with distinctly different base pair specificities were compared in their effects on the formation and elongation of the transcription complex from the lac UV5 promoter in a cell-free system. All were tested at drug levels which killed 90% of cells in a clonogenic survival assay. Cisplatin, a selective alkylator at purine residues, inhibited transcription, decreasing the full-length transcript, and causing blockage at a number of GG or AG sequences, making it probable that intrastrand crosslinks are the blocking lesions. A cyclopropylindoline known to be an A-specific alkylator also inhibited transcription, with blocks at adenines. The aniline mustard chlorambucil, that targets primarily G but also A sequences, was also effective in blocking the formation of full-length transcripts. It produced transcription blocks either at, or one base prior to, AA or GG sequences, suggesting that intrastrand crosslinks could again be involved. The non-alkylating DNA minor groove binder Hoechst 33342 (a bisbenzimidazole) blocked formation of the full-length transcript, but without creating specific blockage sites. A bisbenzimidazole-linked aniline mustard analogue was a more effective transcription inhibitor than either chlorambucil or Hoechst 33342, with different blockage sites occurring immediately as compared with 2 h after incubation. The blockages were either immediately prior to AA or GG residues, or four to five base pairs prior to such sites, a pattern not predicted from in vitro DNA-binding studies. Minor groove DNA-binding ligands are of particular interest as inhibitors of gene expression, since they have the potential ability to bind selectively to long sequences of DNA. The results suggest that the bisbenzimidazole-linked mustard does cause alkylation and transcription blockage at novel DNA sites. in addition to sites characteristic of untargeted mustards.
Antineoplastic Agents, Alkylating↗
Mutations induced by some DNA minor groove binding alkylators in AS52 Chinese hamster cells.
Nitrogen mustards are commonly used in cancer chemotherapy. They interact with DNA at electronegative sites, primarily forming N7 guanine mono-adducts and interstrand cross-links. Targeting nitrogen mustards to DNA by attachment of a DNA minor groove binding carrier such as the bisbenzimidazoles Hoechst 33258 (pibenzimol) or Hoechst 33342 (HOE) makes it possible to direct DNA alkylation to more specific stretches of DNA. We have performed a detailed molecular analysis of 6-thioguanine resistant clones arising in Chinese hamster AS52 cells after treatment with HOE, in comparison with a mono- and bifunctional pair of bisbenzimidazole-targeted nitrogen mustards (MGBs). HOE showed no significant ability to induce 6-thioguanine resistant mutants, possibly because drug-treated cells are highly susceptible to apoptosis within very short times. Neither of the MGBs caused the rapid cell death seen with the bisbenzimidazole. However, both MGBs were weaker mutagens than previously found for undirected mustards in the same system, an effect that we suggest could relate to greater structure-directed binding to less mutable DNA sites in the minor groove. Additionally, the nature of some of the mutants suggested there may be a small component of topo I and/or II-mediated events in the mutagenicity of the MGBs. Both MGBs showed high activity in causing deletion mutations, which may be due to errors in attempted repair of the complex lesions formed by minor groove targeted alkylators.
Alkylating Agents↗
The study of antigenotoxic effects of dietary fibre is lost in a confused concept.
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Animals↗
Comparative effects of three resistant starch preparations on transit time and short-chain fatty acid production in rats.
A high-fiber diet may protect against colon cancer because of the butyrate generated in the colon by bacterial fermentation of nonstarch polysaccharides. Butryrate can reverse neoplastic changes, at least in vitro, and resistant starch (RS) represents a source of butyrate in vivo. We examined the effects of replacing normal maize starch in the diet of rats with three preparations of RS on the amounts of starch, butyrate, and other short-chain fatty acids in the cecum. We examined the effects on fecal bulking and transit time, which have been suggested to protect against colon cancer. The RS preparations that we tested were potato starch, high-amylose maize starch, and an alpha-amylase-treated high-amylose maize starch. All had major effects on fecal weight and on the weight of the cecum but only slightly shortened transit times. All increased the amount of starch reaching the cecum and increased short-chain fatty acid production in the cecum; potato starch had the greatest effect and high-amylose maize starch the least. Potato starch, unlike high-amylose maize starch, enhanced the proportion of butyrate. Thus there were marked differences among sources of RS, even though these were all classified as RS2. The significance for colon cancer is discussed.
Animals↗