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Biomedical subjects

M Fenech

Publications and source records attributed to M Fenech.

At least 55 records · Page 3Linked to original sources
PubMed1995

A comparison of lymphocyte micronuclei and plasma micronutrients in vegetarians and non-vegetarians.

We performed a biochemical and cytogenetic epidemiological study to establish if there are significant differences between vegetarians (V) and non-vegetarians (NV) in their peripheral blood lymphocyte micronucleus (MN) index, which is a measure of chromosome damage rate. The levels of plasma vitamin C (VIT-C), vitamin E (VIT-E), vitamin B12 (B12) and folic acid were also analysed to assess if differences in chromosome damage rates were associated with these potentially antimutagenic micronutrients. Volunteers were classified as either 'vegetarian' if they had abstained from eating any flesh foods for at least 3 years prior to the study or 'non-vegetarian' if they consumed meat or meat products at least 5 days/week for at least 3 years before participation in the study. The volunteers in the study consisted of 47 male and 79 female V and 66 male and 72 female NV, all of whom were non-smokers for at least 3 years prior to the study. The age of the volunteers varied between 20 and 89 years. There was no significant difference in the slope of the age-related increase in MN index of V and NV of either sex. However, the MN index was significantly lower in NV males in the age group 20-40 years and significantly lower for V males in the 41-60 years age group. No difference between the MN index of older males was detectable and there also was no difference in the MN index of V and NV females across all age groups. V were generally found to have significantly higher plasma levels of VIT-C and folic acid, significantly lower levels of B12, and similar levels of VIT-E when compared with NV. VIT-C correlated positively with MN index in young males, but the reverse was true for B12. In young females folate and B12 appeared to correlate negatively with MN index. VIT-E had no apparent impact on MN index. These data suggest that the level of folate and B12 may be more important than VIT-C or VIT-E in minimizing chromosome damage rates in human lymphocytes. Overall, the data from this study do not support the hypothesis that V have a lower genetic damage rate than NV.

Adult↗
PubMed1994

Micronucleus induction in cytokinesis-blocked mouse bone marrow cells in vitro following in vivo exposure to X-irradiation and cyclophosphamide.

A cytokinesis-block micronucleus (MN) method for the simultaneous but separate measurement of chromosome damage in erythroid and myeloid bone marrow cells is described. MN induction in cytokinesis-blocked mouse bone marrow cells in vitro following in vivo exposure to x-ray or cyclophosphamide (CP) was investigated. Immediately after whole body irradiation with acute doses of either 0, 1, 2 or 4 Gy x-rays, or 2 hr after treatment with either 0, 12.5, 25, or 50 mg CP/kg body weight, bone marrow cells were collected and then cultured in medium supplemented with 3.0 micrograms/ml cytochalasin B for 24 hr. The binucleated cells were scored in erythroid, myeloid, lymphoid and other cells. The myeloid/erythroid (M/E) ratio was decreased by x-irradiation or CP treatment in a dose-dependent manner. The dividing index (DI; binucleated cells/binucleated + mononucleated cells; %) was decreased in both erythroid and myeloid cells in the same manner. Dose-dependent increases in MN frequency were observed following x-irradiation in both erythroid and myeloid cells. A similar dose-dependent MN induction was observed with CP. The MN frequency in myeloid cells was much greater than in erythroid cells (about 4-fold following 4 Gy exposure, and more than 10-fold after 50 mg/kg CP). Lymphoid and other cells were not suitable for scoring DI and MN frequency because of insufficient numbers of binucleated cells. These results suggest that micronuclei can be identified in both myeloid and erythroid cells and that myeloid cells are more susceptible to x-ray or CP-induced chromosomal damage than erythroid cells as expressed by MN induction.

Animals↗
PubMed1994

The relationship between micronuclei in human lymphocytes and plasma levels of vitamin C, vitamin E, vitamin B12 and folic acid.

The cytokinesis-block micronucleus assay is increasingly being applied to the study of spontaneous or induced genetic damage in human lymphocytes, but little is known about dietary and other lifestyle factors that could influence this index. As part of a larger study investigating the role of dietary factors on baseline genetic damage in human lymphocytes from 152 non-smoking females and 113 non-smoking males evenly distributed between the ages of 20 and 87 years, we have measured (a) the micronucleus (MN) frequency and (b) the plasma level of the anti-oxidant vitamins C and E and the B vitamins folic acid and B12. Multiple regression analysis indicated that age (beta value = 0.598, P < 0.0001) was the most important factor influencing the variance of micronucleus frequency in females, while micronutrient levels had no apparent significant effects on genetic damage. In males age was also the predominant factor (beta value = 0.505, P < 0.0001) influencing genetic damage, but vitamin-C level also contributed positively and significantly to the observed MN frequency (beta value = 0.220, P < 0.0228). To avoid the potential confounding effect of collinearity between variables we also performed separate simple regression analysis for each plasma micronutrient in relation to age-adjusted micronucleus frequency; the results from this analysis again showed a significant and positive effect of plasma vitamin C on age-adjusted micronucleus frequency in males only (beta value = 0.188, P = 0.0503), while no effect was observed for the other micronutrients in both sexes. In view of the predominant age effect, we also focused on the data obtained in the youngest age groups of both sexes. In view of the predominant age effect, we also focused on the data obtained in the youngest age groups of both sexes (i.e. 20-30 years olds) and found (a) that the MN frequency in young males is significantly and positively correlated with plasma vitamin C levels (r = 0.823, P < 0.001) but negatively correlated with plasma vitamin B12 status (r = -0.799, P < 0.001) and (b) in females the only significant correlation was an inverse relationship between MN frequency and the combined folate and vitamin B12 plasma levels (r = -0.4632, P < 0.030).(ABSTRACT TRUNCATED AT 400 WORDS)

Adult↗
PubMed1994

The origin of micronuclei induced by cytosine arabinoside and its synergistic interaction with hydroxyurea in human lymphocytes.

Using human lymphocytes and the cytokinesis-block micronucleus (CBMN) assay we have recently shown that excision-repairable DNA lesions induced by methylnitrosourea (MNU) and ultraviolet (UV) light (254 nm) can be converted to micronuclei (MNi) within one cell cycle if cytosine arabinoside (ARA-C) is added during the G1 phase of the cell cycle after mitogen stimulation. We have proposed that this conversion resulted from the inhibition by ARA-C of the gap-filling step during excision repair which results in the formation of single-stranded breaks at repair sites; these breaks are then converted to chromatid or chromosome breaks and subsequently to MNi on completion of nuclear division. To confirm this hypothesis we have examined the origin of MNi induced by ARA-C by using anti-kinetochore antibodies and found that between 77 and 86% of these MNi are kinetochore-negative which supports the idea that they originate mainly from acentric chromosome fragments. We have also demonstrated that combining ARA-C treatment with hydroxyurea (HU) during G1 provides a synergistic improvement in the conversion of spontaneous (4-fold increment) or MNU-induced (2-fold increment) excision-repairable DNA lesions to MNi when compared to the effects with ARA-C alone. HU treatment on its own did not influence micronucleus expression in untreated cells and cells treated with MNU.

Adult↗
PubMed1993

Mouse and human micronucleus models for assessing genotoxicity of whole foods in intervention studies.

There is an emerging need to apply classical genotoxicity assays to assess, in vivo, the carcinogenic or anti-carcinogenic potential of traditional and novel foods. The micronucleus techniques in peripheral blood erythrocytes and lymphocytes are two of the methods that have been used for assessing chromosome-damaging effects following and during dietary interventions with whole foods. The advantage of this approach stems from the practicality of the methods and the possibility of using these techniques in both human and rodent models. The potential application of these methods is illustrated by comparison of recent examples in the literature and from ongoing research.

Animals↗
PubMed1993

The cytokinesis-block micronucleus technique: a detailed description of the method and its application to genotoxicity studies in human populations.

The development of the cytokinesis-block (CB) technique has transformed the human-lymphocyte micronucleus assay (MN) into a reliable and precise method for assessing chromosome damage. Recent studies in our laboratory have confirmed that this method is a sensitive indicator of in vivo radiation exposure in (a) patients undergoing fractionated partial-body radiotherapy and (b) rodents exposed to uniform whole-body irradiation, thus supporting the application of the cytokinesis-block micronucleus (CBMN) assay for biological dosimetry. To further define the use of this assay in biomonitoring we performed extensive studies to determine the spontaneous level of MN in normal human populations and its relationship to various life-style factors. We have also developed a new variation to the CBMN assay that permits the conversion of excision-repairable lesions to MN within one cell-cycle using cytosine arabinoside. With this method the slope of the in vitro dose-response curves was increased by a factor of 1.8 for X-rays, 10.3 for ultraviolet (UV, 254 nm) radiation and approximately 40-fold for methylnitrosourea. Consequently the CBMN assay can now be used to measure not only whole chromosome loss or chromosome breaks but also excision-repair events. The versatility and simplicity of the CBMN assay together with new developments in automation should ensure its successful application in monitoring exposed populations as well as in identifying mutagen-sensitive individuals within a population.

Animals↗
PubMed1993

Effect of cooked meat on micronucleus frequency.

The possibility that inclusion of cooked meat in the diet could elevate chromosome damage risk was investigated in mice fed either a cereal diet only or a cereal diet supplemented with mutton cooked by microwaving, pan-frying or charcoal-barbecueing. The meat was provided as a supplement to the standard cereal-based diet on an approximately one-to-one basis. Chromosome damage indices included the cytokinesis-block micronucleus assay in spleen lymphocytes, the bone marrow micronucleus assay in erythrocytes and two versions of the colonic micronucleus assay. These indices were assessed after 3 or 6 months of feeding, and under each circumstance no increment in the micronucleus frequency in any of the tissues could be detected in the groups fed cooked meat when compared with each other or with the group fed the cereal diet only. There was also no correlation between the micronucleus index in lymphocytes and body weight. These data suggest that, over the time-frame studied, inclusion of meat in the diet and the extent to which meat is cooked are unlikely to be important variables affecting the micronucleus index.

Animals↗
PubMed1993

The cytokinesis-block micronucleus technique and its application to genotoxicity studies in human populations.

The development of the cytokinesis-block (CB) technique has made the human lymphocyte micronucleus assay (MN) a reliable and precise method for assessing chromosome damage. Recent studies in our laboratory have confirmed that this method is a sensitive indicator of in vivo radiation exposure in patients undergoing fractionated partial-body radiotherapy and rodents exposed to uniform whole-body irradiation, thus supporting the application of the cytokinesis-block micronucleus (CBMN) assay for biological dosimetry. To further define the use of this assay in biomonitoring, we have also undertaken extensive studies to determine the spontaneous level of MN in normal human populations and its relationship to various lifestyle factors. During the past year, we have also developed a new variation to the CBMN assay that enables the conversion of excision-repairable lesions to MN within one cell-cycle using cytosine arabinoside. With this method the slope of the in vitro dose-response curves was increased by a factor of 1.8 for X-rays, 10.3 for ultraviolet (254 nm) radiation, and approximately 40-fold for methylnitrosourea. Consequently, the CBMN assay can now be used not only to measure whole chromosome loss or chromosome breaks but also excision repair events. The versatility and simplicity of the CBMN assay together with new developments in automation should enable its successful application in monitoring exposed populations as well as identifying mutagen-sensitive individuals within a population.

Cell Cycle↗
PubMed1992

Conversion of excision-repairable DNA lesions to micronuclei within one cell cycle in human lymphocytes.

The human lymphocyte micronucleus (MN) assay is relatively insensitive to genotoxic agents that predominantly induce excision-repairable lesions such as adducts and abasic sites. In this study we have explored the possibility of using cytosine arabinoside (ARA) to convert excision-repairable DNA lesions to micronuclei (MN) within one cell cycle. The system consisted of human lymphocytes as target cells, the cytokinesis-block (CB) method for identifying cells that had completed one nuclear division only, and X-rays, methylnitrosourea (MNU), and ultraviolet light (UV) as mutagens. With each mutagen we have observed significant increments in induced MN in the cultures that had also been treated with ARA during G1. The slope of the dose-response curves for induction of MN was increased by a factor of approximately 1.8 for X-rays and 10.3 for UV and significant MNU induction of MN was only achieved in the cultures treated with ARA. Furthermore, a 24-hr gap between mutagen exposure and the start of the assay did not abolish the increased sensitivity in the cultures treated with ARA. These observations suggested that the combined ARA and cytokinesis-block micronucleus (CBMN) method may enhance the detection of exposure to genotoxic agents that predominantly induce excision-repairable lesions.

Adult↗
PubMed1992

Micronucleus induction in bone-marrow cells following consumption of cooked beef in mice. Preliminary investigations.

The bone-marrow micronucleus assay was used to investigate whether the consumption of cooked meat could induce chromosome damage. There was no difference in the micronucleus frequency of mice on normal diet (cereal-based, non-purified diet) and mice on normal diet supplemented with microwaved meat. However, supplementing the normal diet with well-done pan-fried meat or rare charcoal-barbecued meat or well-done charcoal-barbecued meat produced significant increments in the micronucleus frequency of polychromatic erythrocytes--the increments were of the order of 73% (p = 0.045), 90% (p = 0.047) and 136% (p = 0.001) respectively and they were observed after a 21-day feeding trial. These results suggest that ingestion of well-done pan-fried or barbecued meat may increase genetic damage, however, the accompanying decreased intake of vegetable constituents may have also contributed to the observed changes.

Animals↗
PubMed1991

Cloning and characterization of the rad4 gene of Schizosaccharomyces pombe; a gene showing short regions of sequence similarity to the human XRCC1 gene.

The rad4.116 mutant of the fission yeast Schizosaccharomyces pombe is temperature-sensitive for growth, as well as being sensitive to the killing actions of both ultraviolet light and ionizing radiation. We have cloned the rad4 gene by complementation of the temperature sensitive phenotype of the rad4.116 mutant with a S. pombe gene bank. The rad4 gene fully complemented the UV sensitivity of the rad4.116 mutant. The gene is predicted to encode a protein of 579 amino acids with a basic tail, a possible zinc finger and a nuclear location signal. The amino terminal part of the predicted rad4 ORF contains two short regions of similarity to the C-terminal part of the human XRCC1 gene. Codon usage suggests that the gene is very poorly expressed, and this was confirmed by RNA studies. Gene disruption showed that the rad4 gene was essential for the mitotic growth of S. pombe.

Base Sequence↗
PubMed1990

Micronuclei in cytokinesis-blocked lymphocytes of cancer patients following fractionated partial-body radiotherapy.

We applied the cytokinesis-block micronucleus assay to measure chromosome damage in lymphocytes of 11 cancer patients undergoing fractionated partial-body irradiation. Measurements performed before, during and after cessation of radiotherapy showed a dose-related increase in micronucleus frequency in each of the patients studied. When the results for micronucleus frequency (Y) were plotted against the estimated equivalent whole-body dose (X) the dose-response relationship obtained was Y = 75.8X + 49.5 (r = 0.783, P less than 0.0001). A general decline in MN frequency was observed during the post-treatment period down to 57 per cent (+/- 10) after 12 months but there was considerable variation between individuals. The advantages and disadvantages of the application of the cytokinesis-block micronucleus assay as a biological dosimeter for lymphocytes irradiated in vivo are discussed.

Adult↗
PubMed1989

Kinetochore detection in micronuclei: an alternative method for measuring chromosome loss.

Whole chromosomes within micronuclei (MN) in cytokinesis-blocked human lymphocytes were detected by using anti-kinetochore antibodies obtained from the serum of scleroderma patients. The primary antibody was localized using a peroxidase-labelled second antibody followed by a nickel chloride modification of the diamino-benzidine reaction to give a permanent slide preparation. Between 82 and 92% of colchicine-induced MN were shown to be kinetochore-positive. Results for spontaneously occurring micronuclei in young (20-35 years) and elderly (greater than 65 years) subjects indicated that 42 (+/- 6) and 50 (+/- 6)%, respectively, contained kinetochores. Perhaps the more novel observation was that approximately 12% of X-ray-induced micronuclei were kinetochore-positive and thus could have been the result of whole chromosome loss events. Kinetochore-detection in micronuclei provides a new approach to measure chromosome loss and therefore may be important in identifying aneuploidy-inducing agents.

Adult↗
PubMed1988

Preliminary studies on scoring micronuclei by computerised image analysis.

Initial studies of the use of computerised image analysis to determine micronucleus frequencies in human lymphocytes that have completed one nuclear division are described. Two methods, based on (a) bromodeoxyuridine incorporation and (b) cytokinesis blocking with cytochalasin-B, were studied. The former method is directly amenable to automation. Cytokinesis-blocked cells could not be automatically recognised by image analysis but it was possible to obtain the correct micronucleus frequency from the integrated optical density histograms by using the mononucleate/binucleate cell ratio obtained by visual analysis. The mean (+/- 1 S.E.) integrated optical density of X-ray-induced micronuclei was 11.2% (+/- 1.1) of that measured for nuclei of G1 cells.

Automation↗
PubMed1988

Activation of the food-derived mutagen 2-amino-3-methylimidazo[4, 5-f]quinoline by human-liver microsomes.

The ability of human-liver microsomes to metabolically activate the food-derived heterocyclic amine, 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), and the model mutagen, 2-aminofluorene (AF), has been investigated using Salmonella typhimurium TA98. In 6 subjects tested the number of revertants produced by 0.1 micrograms IQ per mg microsomal protein varied from 11, 830 +/- 320 to 42, 830 +/- 290 (mean +/- SD). With the same livers and a dose of 10 micrograms AF per plate the number of revertants varied from 15,770 +/- 1600 to 29,380 +/- 810 per mg microsomal protein. Metyrapone and alpha-naphthoflavone caused differential inhibition of the mutagenesis of both IQ and AF indicating the involvement of different forms of cytochrome P450 in the metabolic activation of these amines in human-liver microsomes. In presence of human-liver microsomes IQ produced no detectable increase in mutations at the hypoxanthine phosphoribosyl transferase locus in lymphocytes and caused no increase in micronuclei formation at realistic exposure levels.

Benzoflavones↗