Biomedical subjects

K Alexandrov

Publications and source records attributed to K Alexandrov.

At least 55 records · Page 3Linked to original sources

PubMed1992

Expression of pulmonary cytochrome P4501A1 and carcinogen DNA adduct formation in high risk subjects for tobacco-related lung cancer.

Cigarette smoking is the strongest risk factor for lung cancer (LC), but genetically determined variations in pulmonary metabolism of tobacco-derived carcinogens may affect individual risk. Results from a case-control study on LC patients demonstrated the pronounced effect of tobacco smoke on pulmonary xenobiotic metabolism and prooxidant state, and suggested the existence of a metabolic phenotype at higher risk for tobacco-associated LC: LC patients who were recent smokers had significantly induced BP-3-hydroxylase (AHH) and ethoxycoumarin O-deethylase (ECDE) activities in lung parenchyma, when compared with smoking non-cancer patients. In recent smokers, lung AHH activity was positively correlated with the level of tobacco smoke-derived DNA adducts as determined by 32P-postlabelling. Pulmonary AHH activity also showed a good correlation with the intensity of immunohistochemical staining for cyt. P4501A by a monoclonal Ab in lung tissue sections: smoking and peripheral type of lung cancers were positively related to high levels of this cyt. P450 species, probably reflecting high rates of induction. These results suggest that high pulmonary CYP1A1 expression (controlling in part carcinogen DNA-adduct formation) in tobacco smokers, appears to be associated with LC risk. High risk subjects may thus be identifiable through genotyping assays for CYP1A1 polymorphism.

Aryl Hydrocarbon Hydroxylases↗

PubMed1992

Stereoselective metabolism of (-)-benzo[a]pyrene-7,8-diol by human lung microsomes and peripheral blood lymphocytes: effect of smoking.

Benzo[a]pyrene (B[a]P)-tetrols formed after stereoselective cytochrome P450-dependent metabolism from (-)-trans-7,8-dihydroxy-7,8- dihydrobenzo[a]pyrene [(-)-B[a]P-7, 8-diol] by lung microsomes (n = 19) and peripheral blood lymphocytes (n = 13) from lung cancer patients were measured, and the effect of smoking explored. B[a]P-tetrols were quantified by an HPLC/fluorescence assay with a detection limit of approximately 300 attomol, after incubation with peripheral blood lymphocytes or microsomes from lung cancer patients who were current cigarette smokers, ex-smokers and non-smokers. In lymphocytes from these subjects, high, medium and low metabolic activities respectively for (-)-B[a]P-7,8-diol to tetrol conversion were found, but there was no statistically significant difference between smokers, ex-smokers and non-smokers. When the B[a]P-tetrol formation by human lung microsomes was measured, recent smokers had 4- to 7-fold higher (P = 0.04) metabolic activity than ex-smokers and non-smokers. The mean lung microsomal arylhydrocarbon hydroxylase (AHH) activity was three times higher in smokers than in non-smokers and was undetectable in ex-smokers. AHH activity was correlated with tetrol formation in the same lung microsomal samples (r = 0.62, P less than 0.01 in smokers; and r = 0.67, P less than 0.01 in all subjects). When subjects were grouped according to smoking habits, however, no correlation was detected between mean tetrol formation by lung microsomes and that of lymphocytes. Thus, lymphocytes cannot serve as a surrogate for lung microsomes concerning the pulmonary metabolism of (-)-B[a]P-7,8-diol. The much higher B[a]P-tetrol formation observed in lung microsomes from smokers is in accord with a reported higher pulmonary AHH activity, cytochrome P450IA level, and CYP1A1 gene expression in recent tobacco smokers.

Adolescent↗

PubMed1992

Carcinogen metabolism in human lung tissues and the effect of tobacco smoking: results from a case--control multicenter study on lung cancer patients.

Cigarette smoking is the strongest risk factor for lung cancer, but genetically determined variations in the activities of pulmonary enzyme that metabolize tobacco-derived carcinogens may affect individual risk. To investigate whether these enzymes (e.g., CYP1A-related) can serve as markers for carcinogen-DNA damage, lung tissue specimens were taken during surgery from middle-aged men with either lung cancer or non-neoplastic lung disease. Phase I [aryl hydrocarbon hydroxylase (AHH), ethoxycoumarin O-deethylase (ECOD)] and phase II (epoxide hydrolase, UDP-glucuronosyltransferase, glutathione S-transferase) enzyme activities, glutathione and malondialdehyde contents were determined in lung parenchyma and/or bronchial tissues; some samples were also analyzed for DNA adducts, using 32P-postlabeling. The data were then analyzed for the following: a) differences in metabolic profiles between bronchial and parenchymal lung tissue; b) the effect of recent exposure to tobacco smoke on enzyme inducibility and benzo[a]pyrene metabolism; c) differences in enzyme inducibility between lung cancer and non-lung cancer patients; d) the effect of smoking on metabolism of mutagens in vitro; e) pulmonary DNA adduct levels and AHH activity in lung parenchyma of smokers and ex-smokers; f) lipid peroxidation products in lung tissue from lung cancer and non-lung cancer patients, as related to smoking habits and degree of airway obstruction; and g) prognostic value of AHH pulmonary activity in lung cancer patients. The results demonstrate a pronounced effect of tobacco smoke on pulmonary metabolism of xenobiotics and prooxidant state and suggest the existence of a metabolic phenotype at higher risk for tobacco-associated lung cancer.

7-Alkoxycoumarin O-Dealkylase↗

PubMed1990

A new sensitive fluorometric assay for the metabolism of (--)-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene by human hair follicles.

A new sensitive fluorometric assay was established to measure the stereospecific formation of benzo[alpha]pyrene tetrols formed after cytochrome P450-dependent metabolism of (--)-7,8-dihydroxy-7,8- dihydrobenzo[a]pyrene by human hair follicles. This simple assay requires three human hair follicles and a low (0.5-2.0 microM) substrate concentration and has a limit of detection of approximately 0.3 fmol of tetrols. Freshly isolated human hair follicles from 20 adult volunteers (10 non-smokers and 10 smokers) were assayed. While intersubject and seasonal variations were observed, the assay was found to be reproducible for a given subject. This rapid and non-invasive assay provides a new means for metabolic phenotyping of human subjects for their capacity to metabolize (--)-7,8-dihydroxy-7,8-dihydrobenzo[alpha]pyrene to its carcinogenic form (+)anti-benzo[a]pyrene diolepoxide.

Adult↗

PubMed1990

In vivo DNA adduct formation by benzo(a)pyrene in mouse and rat epidermal and dermal fibroblasts after topical application of an initiating dose of benzo(a)pyrene.

In vivo adduct formation by benzo[a]pyrene (BP) has been compared in mouse and rat epidermal keratinocytes and dermal fibroblasts after topical application of an initiating dose of carcinogen. The BP-DNA adducts were analyzed by chromatography and acid hydrolysis of BP-deoxyribonucleoside adducts to BP-tetrols. BP was dissolved in acetone and applied, at similar doses per unit area (100 nmol/mouse and 240 nmol/rat), to 50-day-old Swiss mice and 35-day-old Wistar rats. Epidermal and dermal cells were isolated twenty four hours later. Reverse-phase HPLC of BP-deoxyribonucleoside adducts demonstrated the presence of three BP-deoxyribonucleosides adducts in mouse epidermal cells and one in mouse dermal cells. An unknown product (0.13 and 0.04 pmol/mg mouse epidermal and dermal cell DNA respectively) eluted before the BP-7,10/8,9-tetrol marker, at same relative position as 9-OH-BP-DNA adduct. The major adduct formed in mouse epidermal keratinocytes and dermal fibroblasts was dGuo modified by (+)-anti-BPDE and accounted for more than 70% of the adducts. Acid hydrolysis of the individual BP-DNA adducts was used to identify the BP-DNA adducts formed in mouse epidermal and dermal cells as anti- and syn-BPDE-dGuo. Twenty four hours after topical application of BP, the total levels of modified deoxyribonucleosides and (+)-BPDE-dGuo were 3 times greater in mouse epidermal cells than in dermal cells. The ratios of anti-BPDE to syn-BPDE was 17:1 and 12:1 in mouse epidermal and dermal cells DNA, respectively. This work provides the evidence that, at an initiating dose, 3H modified deoxyribonucleosides of rat epidermal keratinocytes and dermal fibroblasts are not detectable. This may be essential for the resistance of rat skin to the carcinogenic action of benzo[a]pyrene.

Administration, Topical↗

PubMed1989

Evidence for substantial formation of r-7,t-8-dihydroxy-c-9,10-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene- deoxyguanosine in human lymphocytes treated in vitro with benzo[a]pyrene.

The possibility that the amounts of r-7,t-8-dihydroxy-t-9,10-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene- deoxyguanosine (anti-BaP diol epoxide-dGuo) and r-7,t-8-dihydroxy-c-9,10-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene- deoxyguanosine (syn-BaP diol epoxide-dGuo) may vary in human lymphocyte cultures from different donors was investigated by comparing DNA adducts formed after treatment with [G-3H]benzo[a]-pyrene (4 microM) for 24 h. In most cases, greater than 50% of the DNA adducts were derived from r-7,t-8-dihydroxy-c-9,10-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene (syn-BaP diol epoxide).

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

PubMed1988

Differences in the DNA adducts formed in cultured rabbit and rat dermal fibroblasts by benzo(a)pyrene and (-)benzo(a)pyrene-7,8-diol.

Benzo(a)pyrene (BaP) is highly carcinogenic in rats but is without effect in rabbits when administered s.c. The possibility that BaP-DNA adducts could be responsible for this species difference was investigated by comparing BaP-deoxyribonucleoside adducts formed in dermal fibroblast cultures from Wistar rats and New Zealand rabbits. Treatment with [G-3H]BaP (1.2 microM) for 6, 24, and 48 h produced an essentially qualitative species-specific difference. Over 95% of the DNA adducts in the rabbit dermal cell cultures were derived from anti-BaPDE; the major BaP adduct formed (90%) was (+)-anti-BaPDE-deoxyguanosine. This adduct was formed at very low levels in the rat dermal fibroblasts (7%). These cells contained a large proportion of (+/-)-r-7,t-8-dihydroxy-c-9,10-oxy-7,8,9,10-tetrahydrobenzo(a)pyrene (syn-BaPDE)-DNA adducts (45%) and over 48% of other, unidentified, BaP-DNA adducts. Cells treated with (-)-BaP-7,8-diol (1.2 microM) produced almost exclusively (greater than 99%) (+)-anti-BaPDE-deoxyguanosine in rabbit cells, while the rat cells did not form this product. These results suggest that adducts other than anti-BaPDE-deoxyguanosine may be involved in rat s.c. BaP carcinogenesis; the preferential formation of (+)-anti-BaPDE-deoxyguanosine by rabbit dermal fibroblasts does not directly correlate with the resistance of rabbit dermis to tumor formation.

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

PubMed1988

Comparison of DNA adducts in mouse and rat epidermis versus dermis after topical application of (+/-)r-7,t-8-dihydroxy-t-9,10-oxy-7,8,9,10-tetrahydrobenzo(a)pyrene and (+/-) benzo(a)pyrene-4,5-oxide.

Mice and rats were treated topically with the BP metabolites, anti-BPDE and BPO. Rat epidermal DNA was extensively modified by BPO, while mouse epidermal DNA was preferentially modified by anti-BPDE. Anti-BPDE-dGuo adducts were observed only in mouse dermal DNA. DNA adducts were absent from the rat dermis. This adduct formation could be the reason for the very different in vivo biological effects of BP metabolites in the two species.

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

PubMed1986

Comparison of benzo[a]pyrene-DNA adduct levels in mouse and rat epidermis and dermis.

Male Swiss mice and Wistar rats were treated topically with 250 nmol/mouse and 750 nmol/rat of [3H]benzo[a]pyrene ([3H]BaP). The initial level of total BaP--DNA and the individual modified deoxyribonucleoside adducts were similar in the skin epidermis of the two species. The concentration of these adducts was approximately 3 times less in both mouse and rat dermis. The decreased amount of BaP bound to DNA of mouse dermis may be related to the resistance of this tissue to the carcinogenic action of BaP. The ability of both the mouse (susceptible) and rat (resistant) skin to form BaP-bound products similar in nature and ratio in the epidermal and dermal DNA, suggests that other mechanisms are involved in the difference in the biological response of epidermis versus dermis to the carcinogenic effect of BaP.

Animals↗

PubMed1986

In vivo formation and persistence of DNA and protein adducts in mouse and rat skin exposed to (+/-)benzo[a]pyrene-4,5-oxide.

The objective of the present study was to compare DNA and protein adduct formation of benzo[a]pyrene-4,5-oxide (BPO) in vivo and to determine the persistence of the adducts in both mouse and rat epidermis. (+/-)BPO at a dose of 100 nmol/mouse and 200 nmol/rat was topically applied to male Swiss mice and Wistar rats. Three hours after application, there was 3-fold less binding of BPO to mouse epidermal DNA than to rat epidermal DNA; inversely, the amount of BPO bound to mouse skin protein was 3.6 times higher than in rat skin protein. One and three weeks after application of BPO, persistence of 17-20% of the initial amount of BPO-DNA adducts and 2-4% of initial amount of BPO bound to protein was detected in both mouse and rat skin epidermis. H.p.l.c. analysis of the enzymatic hydrolysates of DNA from mouse and rat epidermis 3 h after application of BPO showed five distinct products: one early-eluting, two BPO-deoxyguanosine (dGuo) (ratio 1.5:1) and two BPO-deoxyadenosine (dAdo) adducts (ratio 2:1). The ratio of the total modified dGuo to the total modified dAdo was 2:1. The amount of total BPO-dGuo and BPO-dAdo adducts was 3.5 times greater in rat than in mouse epidermis. Persistence of the major BPO-dAdo adduct was observed in mouse and rat epidermal DNA, and 1 and 3 weeks after topical application of BPO there was a 6-fold greater amount of the persisting BPO-dAdo adduct in rat skin epidermis than in mouse skin epidermis (4.1 and 0.66 pmol/mg DNA, respectively). Minor amounts of the BPO-dGuo were found to persist in rat skin epidermis DNA.

Animals↗

PubMed1986

In vivo formation and persistence of DNA adducts in mouse and rat skin exposed to (+/-)-trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene and (+/-)-7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene.

The in vivo DNA adduct formation of (+/-)-trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene (BPD) and (+/-)-7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (anti-BPDE) were compared and the persistence and disappearance of the adducts in both mouse and rat epidermis determined. BPD (100 nmol/mouse in 150 microliter acetone and 200 nmol/rat in 300 microliter acetone) and anti-BPDE (77 nmol/mouse in 150 microliter tetrahydrofuran and 154 nmol/rat in 300 microliter tetrahydrofuran) were topically applied to 50-day-old male Swiss mice and 35-day-old Wistar rats. To improve the identification of the DNA adducts formed, an acid hydrolysis technique was used to convert the BPD- and anti-BPDE-deoxyribonucleoside adducts formed in mouse and rat skin to BP tetrols. The modified deoxyribonucleosides and BP tetrols obtained by hydrolysis of adducts were isolated by reverse-phase h.p.l.c. At approximately similar doses per unit area of treated skin, the initial total binding of these compounds to epidermal DNA and the level of modified deoxyribonucleosides was approximately 6-fold lower in rat skin epidermis than in mouse skin epidermis. Similar ratios of (+/-)-anti-BPDE-deoxyguanosine (dGuo) to (+/-)-syn-BPDE-dGuo adducts (5.7 and 6.1, determined by h.p.l.c. analysis of BP tetrols obtained by hydrolysis of modified dGuo) were found in both mouse and rat epidermis a short time (6 h) after topical application of (+/-)-trans-BPD. Three hours after topical application of (+/-)-anti-BPDE, the ratios of BP-7,10/8,9-tetrol to 7/8,9,10-tetrol were 9:1 in mouse epidermal DNA and 6:1 in rat epidermal DNA. One and three weeks after application of these two compounds, only (+)-anti-BPDE-dGuo was detected in mouse epidermis; 2 and 0.2% of the initial (+)-anti-BPDE-dGuo level was found to persist in the epidermal DNA from BPD- and anti-BPDE-treated mice respectively. No DNA adducts were detected in rat epidermis 3 weeks after BPD and anti-BPDE treatment. Thus, 3 weeks after topical application of BPD and anti-BPDE to mouse and rat skin, the DNA adducts completely disappeared from rat epidermis while they persisted in mouse epidermis. The results suggest that: the persistence of (+)-anti-BPDE-dGuo may be related to carcinogenesis in mouse epidermis by BPD and anti-BPDE; the complete disappearance of the anti-BPDE-dGuo adduct may also account in part for the relative resistance of tissue from this species to the carcinogenic action of benzo[a]pyrene.

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

PubMed1984

The lack of transformation activity of 9-hydroxybenzo[a]pyrene related to the absence of modified deoxyribonucleosides in DNA of C3H/10T1/2 cells.

Sephadex LH-20 chromatography of DNA digests of C3H/10T1/2 cells treated with [3H]9-hydroxybenzo[a]pyrene (9-OH-BaP) and [14C]BaP-7,8-diol showed: (i) the presence only of uncharacterized 3H radioactivity eluted in the early portion of the gradient; [3H]9-OH-BaP-4,5-oxide-modified deoxyribonucleosides were not observed. (ii) The total amount of [14C]-labelled radioactivity corresponded to nucleoside moieties which were modified by anti- and syn-benzo[a]pyrene diol-epoxide. The absence of nucleosides modified by 9-OH-BaP-4,5-oxide in C3H/10T1/2 cells observed in this study may account in part for the relative resistance of these cells to the mutagenic and transforming action of 9-OH-BaP.

Animals↗

PubMed1983

The persistence of benzo[a]pyrene diol-epoxide deoxyguanosine adduct in mouse skin and its disappearance in rat skin.

Male Swiss mice and Wistar rats, susceptible and resistant, respectively, to the carcinogenic effects of benzo[a]pyrene, were treated topically with 250 nmol/mouse and 1000 nmol/rat of tritium labelled benzo[a]pyrene (BaP). The initial formation of BaP diol-epoxide deoxyguanosine adduct was approximately similar in the skin epidermis of the two species. After 3 weeks, the persistence of some 6.5% of initial BaP diol-epoxide deoxyguanosine was observed in mouse skin DNA, while this adduct was completely removed from DNA of rat skin. The total excision of BaP diol-epoxide deoxy-guanosine adduct in rat epidermis may contribute in part for the relative resistance of rat skin to the carcinogenic actions of BaP.

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

PubMed1982

Persistence of benzo[alpha]pyrenediolepoxide DNA adduct in mouse skin.

The covalently bound products of [3H]benzo[alpha]pyrene (BP) were determined in the DNA isolated from the skin of mice 3 weeks after application of the carcinogen. In the relatively rapidly proliferating skin the persistence of BP-DNA adducts was observed, 50% of which were unmodified nucleosides. Small amounts of (7R) BPDE I-dG adduct (5% from the initial formation) were found after 3 weeks. The minor adducts observed at 18 h after treatment were excised.

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

PubMed1981

The binding of benzo[a]pyrene to mouse and rat skin DNA.

The covalently bound products of[3H]benzo[a]pyrene (BP) were determined in the DNA from the skin of mice and rats. While the total binding to DNA of rat skin is 3 times less than the DNA of mouse skin, the quality and the distribution of bound products was similar. The formation of (7R)-N2-[10-(7beta, 8alpha, 9alpha-trihydroxy-7,8,9,10-tetrahydrobenzo[a]pyrene)yl] deoxyadenosine ((7R)BPDE I-dA) adduct was found.

Animals↗

PubMed1981

Formation of covalent deoxyribonucleic acid benzo[a]pyrene 4,5-epoxide adduct in mouse and rat skin.

The covalently bound products of [3H]benzo[a]pyrene (BP) were determined in the DNA from the skin of mice and rats. The quality and the distribution of bound products was similar. The formation of products corresponding to 4,5-dihydro-4,5-epoxy benzo[a]pyrene (BPE) and to a further metabolite of 9-hydroxybenzo[a]pyrene bound to DNA were found in the 2 species at the 24 h end point.

Animals↗

PubMed1981

Presence of benzo[a]pyrene metabolizing activities in isolated rat liver nucleoli.

A benzo[a]pyrene (B[a]P) hydroxylase activity and epoxide hydrolase (EH) activity have been found in rat liver nucleoli obtained from untreated (C) and 3-methylcholanthrene (3-MC) pretreated rats. A comparison of the enzyme activities was made in rat nuclei and nucleoli. Light and electron microscopic observations of nucleolar preparations did not reveal significant contamination either by intact nuclei or by nuclear membranes, and glucose 6-phosphatase, a marker of microsomal activity, was not detected in our nucleolar preparations. NADPH cytochrome c-reductase could be measured in C and 3-MC nuclei and very low but detectable activity was found in the nucleoli. Nucleolar preparations revealed significant hydroxylating activity, which was inducible by 3-MC in nuclei but not in nucleoli. The presence of EH in nucleoli was demonstrated with phenanthrene 9,10-oxide and B[a]P 4,5-oxide, but the nucleolar activities were lower than those obtained using intact nuclei. The addition of 1,1,1-trichloropropene 2,3-oxide completely inhibited EH activity. Furthermore the presence of covalently-bound metabolites of B[a]P formed in isolated nucleoli was demonstrated by cytofluoromicroscopy.

Animals↗

PubMed1981

Metabolism of benzo[a]pyrene by brain microsomes of fetal and adult rats and mice. Induction by 5,6 benzoflavone, comparison with liver and lung microsomal activities.

Using brain, lung and liver microsomes as the enzyme source in in vitro assays, benzo[a]pyrene (B[a]P) metabolism was studied in fetuses and dams of mice (C57B1/6) and rats (WAG). Separation and quantitation of B[a]P metabolites were performed by h.p.l.c. Microsomal preparations were tested for cytochrome P-450 dependent O-dealkylation of 7-ethoxycoumarin and epoxide hydrolase activities. Another parameter measured included the conjugation of 1-chloro-2,4 dinitrobenzene to glutathione by cytosolic glutathione-S-transferase activity. The induction of B[a]P metabolism was studied after treatment of animals with 5,6-benzoflavone (BF). Mixed function oxygenase, epoxide hydrolase and glutathione-S-transferase activities were transplacentally inducible after dams were treated with BF. Metabolic activation of B[a]P by fetal brain microsomes was lower in both species than that by fetal lung and liver microsomes, but it was higher in fetuses than in adults. All metabolites of B[a]P increased after BF treatment; the production of 7,8-dihydro-7,8-dihydroxybenzo[a]pyrene (7,8-dihydrodiol B[a]P) was higher in brain microsomes from BF-treated rats than that in mice. In stimulated rats, the formation of 7,8-dihydrodiol B[a]P by fetal brain microsomes were higher than that by fetal lung microsomes, whereas in mice, the opposite was observed. These data suggest that initiation could occur in utero, and partially explain the species-specific differences in susceptibility to transplacental tumorigenesis by polycyclic aromatic hydrocarbons by differences in biotransformation in the target organ.

Animals↗