Biomedical subjects

V M Lakshmi

Publications and source records attributed to V M Lakshmi.

40 records · Page 3Linked to original sources

PubMed1986

Metabolism of the renal carcinogen FNT by peroxidases.

Formic acid 2-[4-(5-nitro-2-furyl)-2-thiazolyl]-hydrazide (FNT) is a renal carcinogen in the rat. The peroxidative activity of prostaglandin H synthase oxidizes FNT into a reactive intermediate which forms 5-(S)-substituted thioether conjugates with glutathione and N-acetylcysteine. These conjugates are also formed during horseradish peroxidase oxidation of FNT. The conjugate was identified by the combined results of comparative u.v./vis. spectrophotometry, chromatographically-assisted hydrodynamic voltammetry and proton n.m.r. spectroscopy. The relative rate of PHS metabolism of FNT was similar to that observed with benzidine and 5-fold faster than ANFT, its 5-nitrofuro-2-aminothiazole analogue. These results indicate that the pathogenic effects of FNT may be caused by its peroxidative activation and that cellular thiols may attenuate the toxic effects of FNT by conjugate formation.

Acetylcysteine↗

PubMed1986

Metabolism and excretion of nitrofurothiazole bladder carcinogens.

Nitrofurothiazoles such as 2-amino-4-(5-nitro-2-furyl) thiazole (ANFT) and N-formylated ANFT (FANFT) are model compounds used in the study of chemical carcinogenesis. FANFT is a more potent uroepithelial carcinogen than ANFT but previous studies have shown extensive deformylation of FANFT to ANFT in vivo and ANFT to be the putative proximate carcinogen. To investigate this paradox, disposition of radiolabeled FANFT and ANFT was determined in rats prepared for clearance experiments. After 2 hr, 27.3 +/- 2.1% of recovered ANFT was excreted in urine compared to 44.2 +/- 1.8% of FANFT (P less than .001). In addition, approximately 20% of both FANFT and ANFT were excreted in bile after 2 hr. The disposition of nonradiolabeled FANFT and ANFT was also determined. The urinary excretion rate for ANFT with i.v. ANFT administration was 0.9 +/- 0.1 nmol/min. Following i.v. FANFT administration, the urinary excretion rate for ANFT was 49.7 +/- 8.6 nmol/min (P less than .001). The elimination half-lives were 23 +/- 3 and less than 5 min, for ANFT and FANFT, respectively. The differences in renal handling of ANFT and FANFT could not be accounted for by differences in protein binding. Large differences were found in urinary metabolite excretion between FANFT and ANFT administration. These results demonstrate deformylation dependent excretion (renal metabolic/excretory coupling) exists for FANFT resulting in much higher concentrations of ANFT reaching the urinary tract than when ANFT only is administered. Biliary excretion accounts for significant early clearance of both ANFT and FANFT. Renal metabolic/excretory coupling may explain the difference in uroepithelial carcinogenicity between FANFT and ANFT.

Animals↗

PubMedDate not supplied

The role of acetylation in benzidine metabolism and DNA adduct formation in dog and rat liver.

To determine whether benzidine is acetylated in dog, like rat, the metabolism of benzidine was assessed with dog and rat liver slices. Slices were incubated with 0.05 mM [3M]benzidine for 4 h. Media and cellular DNA were analyzed for acetylated benzidine metabolites and adducts. In rat, benzidine was rapidly converted to acetylated metabolites. At 1 h, benzidine, N-acetylbenzidine, and N,N'-diacetylbenzidine represented 5%, 23%, and 54%, respectively, of the total radioactivity in media. Within 2 h, 75% of the radioactivity was N,N'-diacetylbenzidine. In dog, 45% of the radioactivity was present in metabolites more polar than benzidine by 4 h. No N-acetylated metabolites were observed in dog liver slice media. To identify acetylated benzidine DNA adducts, N-(deoxyguanosin-8-yl)-N,N'-diacetylbenzidine was prepared and identified by FAB MS. This nucleoside adduct was used to synthesize N-(deoxyguanosin-8-yl)-N-acetylbenzidine and N'-(deoxyguanosin-8-yl)-N-acetylbenzidine. Nucleoside adducts from slices incubated with [3H]benzidine were analyzed by HPLC. With this method of analysis, the 3H-material did not correlate with the synthetic adduct standards. To improve sensitivity and identify liver adducts, a 32P-postlabeling method was developed. 2'-Deoxyguanosine 3'-monophosphate adduct standards of acetylated benzidine were prepared. 32P-Postlabeling analysis demonstrated that rat liver contained only N'-(3'-monophosphodeoxyguanosine-8-yl)-N-acetylbenzidine after a 1- or 4-h exposure to benzidine. In contrast, no acetylated adducts were detected in dog. Results indicate that dog is a nonacetylator with respect to benzidine. The availability of acetylated benzidine nucleotide standards allowed unambiguous identification of N'-(3'-monophosphodeoxyguanosin-8-yl)-N-acetylbenzidine as the adduct present in rat liver slices. These nucleotide adduct standards will be useful in subsequent studies in animals and human.

Acetylation↗

PubMedDate not supplied

Glucuronidation of N-acetylbenzidine by human liver.

N-Glucuronidation is an important pathway in aromatic amine metabolism. This study assessed N-glucuronidation of N-acetylbenzidine by human liver slices and microsomes. With slices, considerable metabolism of [3H]N-acetylbenzidine (0.2 mM) was observed during a 2-hr incubation. N-Acetylbenzidine N'-glucuronide represented significant metabolism in four different human liver samples (6-33% of the total recovered radioactivity following HPLC). Benzidine (11-43%), benzidine N-glucuronide (8-11%), and N,N'-diacetylbenzidine (0-2%) were also formed. The kinetics of N-acetylbenzidine N'-glucuronide formation were investigated using Triton X-100-pretreated microsomes. Data were best described by a two-component Michaelis-Menten model composed of both high-affinity (low KM) and low-affinity (high KM) UDP-glucuronsyltranosferases. The high- and low-affinity KMs were 0.36 +/- 0.02 and 1.07 +/- 0.12 mM, respectively. To help identify the UDP-glucuronosyltransferases metabolizing N-acetylbenzidine, 23 transferase substrates were tested for their ability to inhibit glucuronidation. At 0.25 mM, bilirubin, estriol, and 17-epiestriol were good inhibitors (< 50% of control). Dose-response inhibition studies with estriol and 4-aminobiphenyl demonstrated that each agent reached a plateau as its concentration was increased. IC50 for estriol and 4-aminobiphenyl was 0.15 +/- 0.03 and 0.57 +/- 0.06 mM, respectively. Complimentary inhibition was observed when these agents were combined at maximal inhibitory concentrations. These results suggest that more than one UDP-glucuronosyltransferase metabolizes N-acetylbenzidine. N-Glucuronidation represents a major pathway for N-acetylbenzidine metabolism in humans.

Aged↗