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Exposure to the carcinogen benzopyrene depletes tissue vitamin A: beta-carotene prevents depletion.

Evidence in humans and laboratory animals supports a cancer-protective effect of vitamin A, but the mechanism remains unclear. While vitamin A deficiency causes squamous metaplasia, and lung cancer patients have lower vitamin A status, their serum vitamin A levels are not indicative of deficiency. We hypothesize that local enzymatic degradation of vitamin A can be induced by exposure to carcinogens such as benzopyrene found in cigarette smoke. This study was designed to determine if benzopyrene exposure depletes tissue vitamin A and whether beta-carotene might prevent the depletion. Weanling male Fischer rats were fed a nutritionally complete purified diet, supplemented with or without benzopyrene at 400 mg/kg feed or beta-carotene at 2 g/kg feed. Vitamin A content of the liver, small intestine, and serum was determined by high-performance liquid chromatography. There was no effect of benzopyrene feeding on serum retinol levels through four weeks. However, there was a decline in tissue retinol in the liver and small intestine by two weeks, with a 30% decline by four weeks (p less than 0.05). In rats fed beta-carotene, there was no effect of benzopyrene on tissue vitamin A level. These results indicate that exposure to benzopyrene induces a local tissue vitamin A depletion despite a vitamin A-sufficient diet and maintenance of serum vitamin A levels. A high intake of beta-carotene prevented the vitamin A depletion effect of benzopyrene exposure. Further studies appear warranted to determine whether some of the adverse effects of environmental carcinogens, as found in cigarette smoke, charcoal-broiled meats, and industrial wastes, might be alleviated by dietary intervention.

Analysis of Variance

Increase of urinary putrescine in 3,4-benzopyrene carcinogenesis and its inhibition by putrescine.

A significant increase in putrescine was noted in the urine of mice with experimental s.c. tumors induced by a single injection of 3,4-benzopyrene solution (2.52 mg of 3,4-benzopyrene in 0.5 ml of tricaprylin). When 10 mg of putrescine were added to the 3,4-benzopyrene solution, the development of tumors was completely inhibited and the increase of urinary putrescine in mice was suppressed simultaneously. Animal weight data of a control group receiving only putrescine indicated that the inhibitory effect of putrescine is not due to its toxicity.

Animals

[Eventual presence of 3-4 benzopyrene in the non-saponificable part of sunflower oil].

Within the limit of the research on the chemical composition of the nonsaponds of the edible oils, under execution in our Institute have been affected some cromatografic gas and spectoskopic analysis (U.V.) on a sample of sunflower seeds oil. Concentrating our research essentially on the determination of the presence of 3-4 benzopyrene in the above oil, we have treated about 8,3 gr. of nonsapond with appropriate chemical methodologies, in order to obtain specific samples for the polycyclic aromatic hydrocarbon analysis. The results of the cromatografic gas and spectoskopic of two fractions, obtained through the process of the preparation of the nonsapond sample, compared with those of a standard solution of 3-4 benzopyrene, exclude the presence of the hydrocarbon in the sunflower oil.

Benzopyrenes

[Survey of contamination of food by 3, 4-benzopyrene. I. Demonstration of a universal method of determination. Application to the determination in complete meals in food emporia].

A versatile spectrofluorimetric method is proposed for the determination of 3,4-benzopyrene in foods and biological material. Recording the emission spectrum allows a reliable identification, and determination at a 0,2 ppb level, with 85 p. 100 recovery and good reproductibility. The time required for analysis is decreased, if compared to existing methods. As a test of its reliability, results of a survey on 3,4-benzopyrene content in canteen meals are presented. The values obtain are very low, generally below 1 ppb; their significance is discussed.

Animals

The effects of benzopyrene and safrole on biphenyl 2-hydroxylase and other drug-metabolizing enzymes.

A study was made of the nature and specificity of the increase in biphenyl 2-hydroxylase activity after preincubation of liver microsomal preparations with various carcinogens in vitro. This enhancement of enzyme activity in vitro was investigated in mouse, hamster and rat, and although the rat appears to be atypical in the variation of the pattern of 2- and 4-hydroxylation with age, similar enhancements were detectable in each species examined. An increase in biphenyl 2-hydroxylase activity was apparent 2h after intraperitoneal administration of safrole or benzopyrene to mature Wistar albino rats and appeared to be similar in nature to that observed after preincubation of liver microsomal preparations with the same chemical in vitro. Investigation of other drug-metabolizing enzyme systems suggests that the enhancing effects of carcinogens in vitro are specific for biphenyl 2-hydroxylase. No correlation between the enhancement of biphenyl 2-hydroxylase and inhibtion of biphenyl 4-hydroxylase was apparent.

Acetone

Prevention of 3,4-benzopyrene carcinogenesis in presence of putrescine.

The 3,4-Benzopyrene (3,4-BP) carcinogenesis can be postponed or even completely inhibited in the presence of Putrescine (P). A single s.c. injection of 2.52 mg 3,4-BP in 0.5 ml tricaprylin on female mice (NMRI-strains, 4--5 weeks old, 20--25 g of body weight) induced locally malignant tumors (sarcomas and carcinomas) up to 97% of the animals treated. (132 mice with tumors from 136 animals treated). Animals injected with 3,4-BP plus 10 mg putrescine showed a considerable reduction of tumor incidence. Only three from 38 mice treated developed tumors (8%). The prevention of tumor could not be further improved with higher putrescine amounts, for example 15 mg and 20 mg P. These concentrations were moreover toxic to the animals. Histologically, the tumors developed in the presence of putrescine were rather poor in malignant cells and mitoses were also rare, in contrast to the usual 3,4-BP tumors which were rich in polymorph cells and mitoses.

Animals

Experimental investigations on the influence upon the chemical carcinogenesis. IInd communication: studies with 3,4-benzopyrene.

After subcutaneous application of 0.5 mg 3,4-benzopyrene (BP) to Sprague-Dawley-rats on the 2nd day of life, 50% of the animals developed local fibrosarcomas after 250 +/- 70 days. Additional treatment with immunostimulating (BCG, albumin, vitamin A-acid) or immunodepressive agents (hydrocortisone, cyclophosphamide, methotrexat) which was started 6 days after birth and maintained throughout life, did not influence carcinogenesis with respect to tumor incidences and induction periods of tumors.

Animals

[Determination of 3, 4-benzopyrene in spiruline algae produced and treated by various procedures].

Spirulina algae grow in highly salted natural lakes. Artificial cultivation in tanks with addition of carbon dioxide (natural gas, burned gases) has been studied in order to improve the biosynthesis. A possible 3, 4-benzopyrene (BaP) contamination must be then considered. Several BaP determinations have been performed in batches of algae from bath origins, prepared following different processes (filtration, spray, cylinder drying). BaP contents are very low (2 to 3 ppb) and comparable between batches.

Benzopyrenes

Sequence specificity in the reaction of benzopyrene diol epoxide with DNA.

Benzopyrene diol epoxide (BPDE; (+)-7R,8S-dihydroxy-9S,10R-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene), the ultimate carcinogen derived from the polycyclic hydrocarbon benzo[a]pyrene, reacts principally with the guanine bases in DNA. Nineteen double stranded, self-complementary oligonucleotides, containing deoxyguanosine in various sequence contexts, were each treated with tritium labelled BPDE. The extent of reaction was determined by releasing the BPDE-guanine adduct with acid, isolating it by chromatography on a reverse-phase column, and estimating it by its radioactivity. Oligonucleotides containing an isolated guanine, such as AAGTACTT, were little affected by BPDE. Reactivity was increased where the guanine was flanked by another guanine on the same strand (e.g. TACCTAGGTA) or on the complementary strand (e.g. TATTCGAATA), and was highest in mixed G-C sequences such as ATCCGGAT. The results should help predict major sites of attack of BPDE on cellular proto-oncogenes.

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