Synthesis and evaluation of new heteroaryl nitrones with spin trap properties.

A new series of heteroaryl nitrones were synthesized and evaluated as free radical traps due to the results showed in our previous report. The physicochemical characterization of these new nitrones by electron spin resonance (ESR) demonstrated their high capability to trap and stabilize different atom centered free radicals generated by the Fenton reaction. Additionally, we intensely studied them in terms of their physicochemical properties. Kinetic studies, including the use of a method based on competition and the hydroxyl adduct decay, gave the corresponding rate constants and half-lives at the physiological pH of these newly synthesized nitrones. New nitrones derived from quinoxaline 1,4-dioxide heterocycles were more suitable than DMPO to trap hydroxyl free radicals with a half-life longer than two hours. We explain some of the results using computational chemistry through density functional theory (DFT).

Stearoyl-CoA desaturase-1, a novel target of omega-3 fatty acids for reducing breast cancer risk in obese postmenopausal women.

BACKGROUND/OBJECTIVES: Conversion of saturated fatty acids to monounsaturated fatty acids by the enzyme stearoyl-Co-A-desaturase (SCD-1) is emerging as a major factor in promoting carcinogenesis including breast cancer. The aim of our study was to explore the regulation of SCD-1 by Raloxifene and omega-3 fatty acids in women at increased risk of breast cancer based on high breast density. SUBJECTS/METHODS: As a reflection of SCD-1 activity, we measured the ratios of palmitoleic acid (C16:1n7) to palmitic acid (C16:0) (SCD-16) and oleic acid (C18:1n9) to steric acid (C18:0) (SCD-18) in plasma samples of postmenopausal women enrolled in our clinical trial (NCT00723398) designed to test the effects of the antiestrogen, Raloxifene and/or the omega-3 preparation Lovaza, on breast density, a validated biomarker of breast cancer risk. RESULTS: We report that Lovaza but not Raloxifene-reduced SCD-16 and SCD-18 for the 2-year duration of the trial. Importantly, decreasing levels of SCD-16 and SCD-18 were associated with a progressive reduction in breast density but only in obese women (body mass index ⩾30). CONCLUSIONS: Body mass index-related factors play an important role in the reduction of breast density and hence breast cancer risk by omega-3 fatty acids. SCD-1 may be a useful biomarker in future clinical trials testing the benefit of nutritional interventions in reducing obesity-associated breast cancer risk.

Genotoxicity and carcinogenicity in rats and mice of 2-amino-3,6-dihydro-3-methyl-7H-imidazolo[4,5-f]quinolin-7- one: an intestinal bacterial metabolite of 2-amino-3-methyl-3H-imidazo[4,5-f]quinoline.

BACKGROUND: Compounds formed on the surface of fried or grilled meat and fish may be associated with increased risk of colon cancer. Normal intestinal bacteria can convert one of these compounds, 2-amino-3-methyl-3H-imidazo[4,5-f]quinoline (IQ), to the 7-hydroxy metabolite, 2-amino-3,6-dihydro-3-methyl-7H-imidazolo[4,5-f]quinolin-7-o ne (7-OHIQ), a direct-acting mutagen. PURPOSE: We studied the genotoxicity and carcinogenicity of 7-OHIQ to determine if it is responsible for the colon-specific activity of IQ. METHODS: The effects of pure, synthetic 7-OHIQ on DNA were evaluated in the Ames Salmonella typhimurium TA98 test, with and without an induced rat liver S9 fraction, and in the Williams DNA repair test using freshly explanted rat hepatocytes. 7-OHIQ was also subjected to an in vivo bioassay for 21 months by long-term intrarectal infusion in male F344 rats, using IQ and N-nitrosomethylurea (NMU) given intrarectally as positive tumor-producing controls. The standard NIH-07 rodent diet was supplemented with 15% corn oil to maximize any effect of the infused materials on the colon. A parallel bioassay involved intraperitoneal injection of 7-OHIQ in newborn mice, followed by dietary administration from week 11 to week 67. Again, IQ and NMU were used as positive controls. RESULTS: We confirmed that 7-OHIQ is a direct-acting mutagen in the Ames test, with added S9 liver fraction giving higher mutagenicity. 7-OHIQ was negative in the Williams test, whereas IQ was positive. 7-OHIQ did not induce colon cancer in rats, and in the newborn mouse test it produced only a low incidence of liver neoplasms. CONCLUSIONS: 7-OHIQ is not genotoxic, for to be so classified it must be definitely positive in both the Ames and Williams tests; moreover, it is not carcinogenic, in marked contrast to IQ and NMU.

Effect of amount and types of dietary fat on intestinal bacterial 7 alpha-dehydroxylase and phosphatidylinositol-specific phospholipase C and colonic mucosal diacylglycerol kinase and PKC activities during stages of colon tumor promotion.

It is evident from many studies that the effect of dietary fat on colon tumor promotion depends not only on the amount of fat but especially on fatty acid composition. Animal model studies have shown that diets which are high in omega-6 fatty acids increase colon tumor promotion, whereas diets rich in omega-3 fatty acids have no such enhancing effect. The mechanisms by which the high fat content of the diet promotes colon carcinogenesis may include the production of secondary bile acids in the colon and the modulation of colonic luminal bacterial 7 alpha-dehydroxylase that is involved in generating secondary bile acids, phosphatidylinositol-specific phospholipase C (PI-PLC), and mucosal PI-PLC, as well as diacylglycerol (DAG) kinase and protein kinase C (PKC). In the present study, we investigated the effect of high-fat diets that are rich in omega-3 and omega-6 fatty acids on cecal bacterial 7 alpha-dehydroxylase and PI-PLC, fecal secondary bile acids, and colonic mucosal DAG kinase and PKC activities during different stages of colon carcinogenesis in male F344 rats. At 5 weeks of age, groups of animals were fed a low-fat diet containing 5% corn oil (LFCO). Beginning at 7 weeks of age, all animals, except those intended as vehicle controls, received azoxymethane (AOM) s.c. once weekly for 2 weeks at a dose rate of 15 mg/kg body weight. Vehicle-treated groups received s.c. injections of normal saline. One day after the second AOM or saline treatment, the experimental groups of animals were transferred to a high-fat diet containing 23.5% corn oil (HFCO) or 20.5% fish oil + 3% corn oil (HFFO). One group continued on the LFCO diet. Animals were sacrificed at weeks 1, 12, and 36 after the AOM or saline treatment. Colonic mucosa were harvested at weeks 1, 12, or 36, and the colonic tumor tissues were examined for PKC and DAG kinase activities. Contents of the cecum were analyzed for bacterial 7 alpha-dehydroxylase and PI-PLC activities. Stool samples collected at week 12 were analyzed for bile acids. High corn oil content of the diet significantly increased the cecal bacterial 7 alpha-dehydroxylase and PI-PLC activities as compared to the diets with high fish oil or low corn oil content. Animals fed the HFCO diet excreted higher levels of secondary bile acids, such as deoxycholic acid and lithocholic acid, than those fed the LFCO or HFFO diets. Carcinogen treatment significantly enhanced the activities of DAG kinase and total membrane PKC activities in colonic mucosa compared to saline treatment in all dietary groups. Animals treated with saline or AOM and fed HFCO showed increased levels of DAG kinase and membrane PKC activities in the colonic mucosa when compared to LFCO and HFFO groups. DAG kinase and membrane PKC activities were higher in colon tumors than in the surrounding colonic mucosa, and also increased levels of these enzyme activities were found in the HFCO diet group. These results indicate that the modifying effect of dietary fat on colonic bacterial enzymes, secondary bile acids, colonic mucosal and tumor DAG kinase, and PKC that may play a role in colon carcinogenesis depends on the types and amount of fat given. The colon tumor-enhancing effect of a HFCO diet in contrast to the high dietary fish oil may be, in part, explained on the basis of its modulating effect on these bacterial and colonic mucosal enzymes and colonic secondary bile acids relevant to colon tumor promotion.

LAS, a novel selective estrogen receptor modulator with chemopreventive and therapeutic activity in the N-nitroso-N-methylurea-induced rat mammary tumor model.

The N-nitroso-N-methylurea-induced rat mammary tumor model was used to conduct two types of studies: a prevention study designed to test the ability of the novel selective estrogen receptor modulator lasofoxifene (LAS) to inhibit the development of mammary tumors, and a treatment study designed to test the inhibitory effect of LAS on the growth of established tumors. The prevention study indicated that LAS markedly delayed the emergence of N-nitroso-N-methylurea-induced tumors to an extent similar to that obtained by the established antiestrogen tamoxifen (TAM). At the highest dose administered, both TAM and LAS reduced tumor incidence by 75% and total tumor number by 90% relative to the controls. LAS also reduced the multiplicity of tumors, i.e., the mean number of tumors per rat, and resulted in substantially smaller total tumor burden. In the treatment study, LAS significantly inhibited tumor growth compared with the controls. In addition, whereas none of the untreated tumors regressed completely over the experimental period, 40% of LAS-treated tumors regressed by >50% at the highest dose (10 mg/kg daily). The results of this study in a rat mammary tumor model indicate that LAS has both chemopreventive and chemotherapeutic effects quantitatively comparable with those of TAM.

Compostability assessment of nano-reinforced poly(lactic acid) films.

Nanomaterials can provide plastics with great advantages on mechanical and active properties (i.e. release and capture of specific substances). Therefore, packaging is expected to become one of the leading applications for these substances by 2020. There are some applications already in the market. Nevertheless, there is still some areas under development. A key issue to be analyzed is the end-of-life of these materials once they become waste, and specifically when nanomaterials are used in biodegradable products. The present study evaluated the disintegration, biodegradability, and ecotoxicity of poly(lactic acid) films reinforced with the three following nanomaterials: (1) montmorillonite modified with an ammonium quaternary salt, (2) calcium carbonate and (3) silicon dioxide. Results on disintegration showed that films completely disintegrated into visually indistinguishable residues after 6-7weeks of incubation in composting environment. Moreover, no differences were observed in the evolution of the bioresidue with respect to color, aspect, and odor in comparison with the control. It was also observed that nanomaterials did not significantly reduce the level of biodegradability of PLA (p>0.05). In fact, biodegradation was higher, without finding significant differences (p>0.05), in all the nano-reinforced samples with respect to PLA after 130days in composting (9.4% in PLA+Nano-SiO2; 34.0% in PLA+Clay1; 48.0% in PLA+Nano-CaCO3). Finally, no significant differences (p>0.05) in ecotoxicity in plants were observed as a result of the incorporation of nanoparticles in the PLA matrix.

The influence of printed electronics on the recyclability of paper: a case study for smart envelopes in courier and postal services.

The aim of this paper is to analyse the effects of the presence of printed electronics on the paper waste streams and specifically on paper recyclability. The analysis is based on a case study focussed on envelopes for postal and courier services provided with these intelligent systems. The smart printed envelope of the study includes a combination of both conventional (thin flexible batteries and resistors) and printed electronic components (conductive track layout based on nanosilver ink). For this purpose, a comparison between envelopes with and without these components (batteries, resistors and conductive track layouts) was carried out through pilot scale paper recycling tests. The generation of rejects during the recycling process as well as the final quality of the recycled paper (mechanical and optical properties) were tested and quantitatively evaluated. The results show that resistors are retained during the screening process in the sieves and consequently they cannot end up in the final screened pulp. Therefore, mechanical and optical properties of the recycled paper are not affected. Nevertheless, inks from the conductive track layouts and batteries were partially dissolved in the process water. These substances were not totally retained in the sieving systems resulting in slight changes in the optical properties of the final recycled paper (variations are 7.2-7.5% in brightness, 8.5-10.7% in whiteness, 1.2-2.2% in L(∗) values, 3.3-3.5% in opacity and 16.1-27% in yellowness). These variations are not in ranges able to cause problems in current paper recycling processes and restrict the use of recycled paper in current applications. Moreover, real impacts on industrial recycling are expected to be even significantly lower since the proportion of paper product with printed circuits in the current paper waste streams are much lower than the ones tested in this work. However, it should be underlined the fact that this situation may change over the next years due to the future developments in printed electronics and the gradual penetration of these types of devices in the market.

Four glycoproteins are expressed in the cat zona pellucida.

The mammalian oocyte is surrounded by a matrix called the zona pellucida (ZP). This envelope participates in processes such as acrosome reaction induction, sperm binding and may be involved in speciation. In cat (Felis catus), this matrix is composed of at least three glycoproteins called ZP2, ZP3, and ZP4. However, recent studies have pointed to the presence of a fourth protein in several mammals (rat, human, hamster or rabbit), meaning that a reevaluation of cat ZP is needed. For this reason, the objective of this research was to analyze the protein composition of cat ZP by means of proteomic analysis. Using ZP from ovaries and oocytes, several peptides corresponding to four proteins were detected, yielding a coverage of 33.17%, 71.50%, 50.23%, and 49.64% for ZP1, ZP2, ZP3, and ZP4, respectively. Moreover, the expression of four genes was confirmed by molecular analysis. Using total RNA isolated from cat ovaries, the complementary deoxyribonucleic acids encoding cat ZP were partially amplified by reverse-transcribed polymerase chain reaction. Furthermore, ZP1 was totally amplified for the first time in this species. As far as we are aware, this is the first study that confirms the presence of four proteins in cat ZP.

Characterization of the profile of neurokinin-2 and neurotensin receptor antagonists in the mouse defense test battery.

Defensive behaviors of lower mammals confronted with a predatory stimulus provide an appropriate laboratory model for investigating behavior relevant to human emotional disorders. The mouse defense test battery (MDTB) has been developed because it combines many of the aspects of defense. Briefly, it consists of five tests either associated with potential threat (contextual defense) or the actual presence of an approaching threat (a rat). These latter focus on changes in flight, risk assessment and defensive threat and attack behaviors. Investigations with anxiolytic compounds have shown that these defense reactions may be used to differentiate between several classes of anxiolytic drugs. Here we used the MDTB to compare the behavioral profile of the benzodiazepine diazepam with that of neuropeptide receptor antagonists which have been shown to be involved in the modulation of stress response, namely the NK(2) receptor antagonists, SR48968 (0.01-1mg/kg) and SR144190 (1-10mg/kg), and the NT(1) receptor antagonist, SR48692 (1-30mg/kg). Results showed that all compounds decreased defensive threat/attack, but only diazepam and, to a lesser extent, SR48692 significantly modified risk assessment or flight. Further, none of the neuropeptide receptor antagonists modified contextual defense. Overall, the behavioral profile displayed by diazepam and these latter compounds in the MDTB are consistent with an anxiolytic-like action. However, our results suggest that, while NK(2) and NT(1) receptor antagonists may have limited efficacy on anxiety-related responses including cognitive aspects (i.e. risk assessment), they may have a potential against some forms of anxiety disorders which involve adaptative responses to extreme stress stimuli (e.g. direct confrontation with the threat stimulus).

Tea, or tea and milk, inhibit mammary gland and colon carcinogenesis in rats.

Research was performed on the effect of tea, or tea and milk, instead of drinking water, in rat models of cancer in the mammary gland or colon. Solutions of 1.25% (w/v) black tea, or 1.85% (v/v) milk in tea were prepared three times per week. SD rats were given tea beginning at 42 days of age; one group was gavaged 5 mg 7,12-dimethylbenz[a]anthracene (DMBA) at 49 days of age; another group received 8.4 mg 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) twice per week beginning at age 49, then 14 mg twice a week for 4 weeks more. The groups on DMBA were killed 33 weeks later, and those on IQ 39 weeks later. Tea decreased the mammary gland tumor multiplicity and volume, and milk and tea had a greater protective action. Male F344 rats were given two doses of 15 mg/kg azoxymethane (AOM) on weeks 6 and 7, and some groups started on tea, or tea and milk at 5 weeks; one group started on tea 2 days after AOM. Foci of aberrant crypts in the colon were decreased, after 9 weeks, in the groups on tea, or tea and milk during AOM administration, but not after AOM. Thus, tea decreases mammary tumor induction, and the production of foci of aberrant crypts in the colon. Milk potentiates these inhibiting effects.

The role of fat and calcium in the production of foci of aberrant crypts in the colon of rats fed 2-amino-1-methyl-6-phenylimidazo[4,5-b]-pyridine.

The modulation by dietary fat levels of intestine carcinogenesis is well documented. New developments suggest that calcium ions may also play a role. A rapid bioassay, the induction of foci of aberrant crypts in the colon, was used to explore the interaction between dietary fat and calcium. Male F344 rats 6 weeks of age were placed on diets containing 5 or 20% corn oil, and 0.04 or 0.32% calcium ion, as calcium lactate. Each dietary group was fed 400 ppm 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhlP), and negative controls received the diets alone. A positive control group was given 2 mg N-nitrosomethylurea (NMU) intrarectally four times in a 2-week period. All rats were killed after 9 weeks. The intestinal tract was rinsed with Krebs-Ringer buffer. After staining a 6-cm segment of the descending colon and rectum with 0.2% methylene blue, foci of aberrant crypts were evaluated microscopically. With PhlP as a carcinogen, the rats on a high-fat, low-calcium level had more foci of aberrant crypts than animals on a low-fat level. With the higher calcium level, there were fewer foci and aberrant crypts, but the effect of fat was still significant. With NMU and a low-calcium level, the effect of fat level was evident. However, with the higher calcium intake, there were considerably more foci of aberrant crypts than on the low-calcium level, and the effect of the dietary fat level was not obvious.(ABSTRACT TRUNCATED AT 250 WORDS)

Kinetics and mechanism of the reaction of a nitroxide radical (tempol) with a phenolic antioxidant.

In the absence of redox-active transition metal ions, the removal of Tempol by Trolox occurs by a simple bimolecular reaction that, most probably, involves a hydrogen transfer from phenol to nitroxide. The specific rate constant of the process is small (0.1 M(-1) s(-1)). Metals can catalyze the process, as evidenced by the decrease in rate observed in the presence of diethylenetriaminepentaacetic acid (DTPA). Furthermore, addition of Fe(II) (20 microM ferrous sulfate and 40 microM EDTA) produces a noticeable increase in the rate of Tempol consumption.

Oral enzyme therapy and experimental rat mammary tumor metastasis.

Although there has been much interest over the years in the medical use of orally administered proteolytic enzymes, there is considerable controversy about their efficacy against advanced stages of cancer. In light of this, the goal of the present study was to assess the inhibitory effects of different doses of an orally administered porcine pancreas preparation on the growth and metastasis of the R13762 transplantable rat mammary tumor. Five groups of 12 F-344 female retired breeders were inoculated orthotopically with a 2mm3 tumor implant and placed on the following diets: (1) AIN-76A diet + 20% porcine pancreas preparation (PPP); (2) AIN-76A + 20% PPP + 10 mg Mg citrate/rat/day; (3) AIN-76A + 2% PPP; (4) AIN-76A + 2% PPP + 10 mg Mg citrate and (5) AIN-76A only (control). Primary tumor development was monitored for 40 days and following sacrifice, lungs were excised, stained and metastatic foci quantitated. Metastatic foci were sorted into 3 groups based on their radii: small (<1mm), medium (1-3mm) and large (>3mm), and volumes calculated. The oral enzyme preparation had no effect on primary tumor growth or on body weight change over the duration of the study. The percent (incidence) of rats with pulmonary metastases among the five groups were not significantly different. However, among the three size categories of pulmonary foci, decreased incidence was found only in the large (>3mm) volume subset of the 2% PPP group supplemented with Mg++. When assessed in terms of mean number of pulmonary foci/rat, the 20% PPP group exhibited the highest and controls the lowest frequency with the important exception of the 2% PPP + Mg++ group (large volume) which exhibited the lowest frequency of all treatment groups. In general, the presence of Mg++ resulted in marked decreases in mean number of pulmonary foci/rat compared to groups fed PPP without the Mg++ supplement. Similar results were obtained when foci were quantitated in terms of metastatic volume rather than frequency. The results of this laboratory animal study suggest that to show effective inhibition of metastatic dissemination of the R13762 tumor by PPP, lower doses of PPP and larger numbers of animals, to account for the high variability in the model, will be required.

Recyclability assessment of nano-reinforced plastic packaging.

Packaging is expected to become the leading application for nano-composites by 2020 due to the great advantages on mechanical and active properties achieved with these substances. As novel materials, and although there are some current applications in the market, there is still unknown areas under development. One key issue to be addressed is to know more about the implications of the nano-composite packaging materials once they become waste. The present study evaluates the extrusion process of four nanomaterials (Layered silicate modified nanoclay (Nanoclay1), Calcium Carbonate (CaCO3), Silver (Ag) and Zinc Oxide (ZnO) as part of different virgin polymer matrices of polyethylene (PE), Polypropylene (PP) and Polyethyleneterephtalate (PET). Thus, the following film plastic materials: (PE-Nanoclay1, PE-CaCO3, PP-Ag, PET-ZnO, PET-Ag, PET-Nanoclay1) have been processed considering different recycling scenarios. Results on recyclability show that for PE and PP, in general terms and except for some minor variations in yellowness index, tensile modulus, tensile strength and tear strength (PE with Nanoclay1, PP with Ag), the introduction of nanomaterial in the recycling streams for plastic films does not affect the final recycled plastic material in terms of mechanical properties and material quality compared to conventional recycled plastic. Regarding PET, results show that the increasing addition of nanomaterial into the recycled PET matrix (especially PET-Ag) could influence important properties of the recycled material, due to a slight degradation of the polymer, such as increasing pinholes, degradation fumes and elongation at break. Moreover, it should be noted that colour deviations were visible in most of the samples (PE, PP and PET) in levels higher than 0.3 units (limit perceivable by the human eye). The acceptance of these changes in the properties of recycled PE, PP and PET will depend on the specific applications considered (e.g. packaging applications are more strict in material quality that urban furniture or construction products).

Feeding thiol-containing compounds, derived from vegetables, fails to inhibit N-methylnitrosourea-induced mammary tumorigenesis.

Various thiol-containing compounds have been shown to inhibit chemically-induced tumors in animal models. Two thiol-containing compounds derived from vegetables, namely 1,2 dithiol-3-thione (DTT) and S-methylmethane thiolsulfonate (MMTS), were tested for their chemopreventive activity in the N-methylnitrosourea (NMU)-induced rat mammary tumor model. Each compound was incorporated into the grain-based Teklad 7001 diet and fed to the rats one week prior to initiation with NMU until termination 18 weeks post NMU. DTT was fed at 166 and 500 ppm and MMTS at 200 and 800 ppm. Neither compound exerted a significant inhibitory effect on any index of tumor development including incidence, total tumor, tumor multiplicity, volume or latency. Serum levels of DTT assessed at termination in the 500 ppm DTT group ranged from 10-30 microg/ml. MMTS was undetectable in serum from either MMTS-fed group. The results of this study, using the direct acting carcinogen, NMU, suggest that the chemopreventive effect of thiol-containing compounds may be confined to animal models using carcinogens that require host activation.

A comparison of methods employed to evaluate antioxidant capabilities.

Three different methodologies frequently employed to evaluate the indexes that report the antioxidant capabilities of pure compounds and/or complex mixtures of antioxidants are applied to a series of mono- and polyphenols, as well as to two wine (red and white) samples. These methodologies are based on the bleaching of a stable radical, the effect of the additive upon luminol chemiluminescence induced by peroxyl radicals, and the effect of the additive upon the bleaching of the fluorescence from a dye molecule. Widely different responses are obtained from the different methodologies. These differences are interpreted in terms of the different factors (stoichiometric factors and/or reactivities) that determines the indexes evaluated by these different methodologies.

Detection of nitrated benzene metabolites in bone marrow of B6C3F1 mice treated with benzene.

Benzene, a constituent of cigarette smoke, is a human leukemogen and induces bone marrow toxicity. The mechanism of benzene-induced toxicity is not well-established. We hypothesized that relatively high levels of nitric oxide formed in bone marrow can react with oxygen and/or superoxide anion that is generated during redox cycling of ring-hydroxylated benzene metabolites to yield peroxynitrite as well as other NO-derived intermediates. Peroxynitrite can either directly damage cellular macromolecules or form nitrated toxic metabolites. Toward this end, we investigated whether nitro derivatives of benzene are formed in bone marrow of mice treated with benzene. First, we have characterized products formed during activation of benzene in Fenton's system in the absence or presence of NO-releasing compound in vitro by GC/MS. The result of above experiment prompted us to determine whether similar products can be formed in vivo. Groups of B6C3F1 male mice, eight weeks of age, were given a single intraperitoneal dose of [14C]benzene (400 mg/kg body wt, 9.7 mCi/mmol) or an equal dose of unlabeled benzene in corn oil, and the mice were killed 0.5 or 1 h posttreatment. The control group received only vehicle injections. Organic solvent extractable metabolites from bone marrow, liver, lungs, and blood of mice treated with [14C]benzene were identified by comparison of their respective retention times under two different HPLC conditions with authentic standard samples. These metabolites were further characterized by comparison of their GC/MS properties to those of reference standards. Nitro metabolites, namely, nitrobenzene, nitrobiphenyl, and nitrophenol isomers, were detected in the bone marrow of the mice 1 h after benzene treatment. Formation of nitro derivatives in other tissues was either not observed or was significantly less than that formed in bone marrow. This study clearly demonstrates that nitric oxide is a contributor to benzene metabolism and can form nitrated derivatives that may, in part, account for bone marrow toxicity.

Effect of black tea on azoxymethane-induced colon cancer.

Two sets of experiments on the role of tea in azoxymethane (AOM) induced colon cancer were performed. The first test involved male F344 rats given 1.25% solutions of black tea beginning at 5 weeks of age and ending at 51 days of age. At 6 and 7 weeks of age, they received 15 mg/kg AOM and were held for 50 weeks. Another group received the AOM dosage at 6 and 7 weeks and were placed on the tea solutions 2 days after the last AOM dosage, at 51 days of age, and held for the 50-week period. The end point was the occurrence and multiplicity of colon cancer, classified as in situ, exophytic, invasive and Peyer's patch carcinomas. Tea failed to affect the incidence and multiplicity of colon cancers when given during or after the AOM administration, but tea after AOM increased the multiplicity of exophytic carcinomas. In a second series of tests, solutions of 0.6, 1.25, 1.75 or 2.5% tea were given, beginning 1 week prior to the two AOM doses and extending for 42 weeks. Also, one group received 1.25% tea and 1.85% whole milk. The incidence of exophytic or invasive colon cancer and tumor multiplicity were similar in all treatment groups, although the incidence of exophytic neoplasms was higher with 2.5% tea. Thus, chronic administration failed to significantly change the incidence and multiplicity of the AOM-induced colon cancers. These findings are accounted for by the underlying mechanism, namely the fact that tea solutions do not alter the amount of cytochrome P-4502E1 required for the metabolic activation of AOM.